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Find the Support You Need for Your All in One:
Contact Brother by Email, Chat or Phone
Get the Latest Drivers, Software Updates, Manuals, FAQs, Videos and More!
Locate a Brother Authorized Service Center to provide support on your Brother products.
High-Speed Monochrome Laser All-in-One for Small to Medium Sized BusinessesThe Brother™ MFC-8910DW monochrome laser all-in-one is a smart, cost efficient choice for your busy office or business. It combines high-speed, reliable printing and copying up to 42ppm, high-quality color scanning, and faxing into one durable design. Its advanced duplex capabilities provide automatic two-sided printing and single-pass, two-sided copying/scanning/faxing. Share this all-in-one with others on your Ethernet or wireless 802.11b/g/n network or print from your mobile device via‡: AirPrint™, Google Cloud Print™, Brother™ iPrint&Scan, Cortado Workplace and Wi-Fi Direct®. A legal-size document glass provides ample room to scan or copy bound materials or use the 50-page capacity automatic document feeder for multi-page documents. Plus, a 1 year on-site limited warranty comes standard for worry free use.
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Assisted Unit Distribution What is Housing Assistance?
The Housing Authority of the County of Dona Ana administers both a public housing and Section 8 housing voucher program. The housing authority owns and manages 1 projects which contain 5 affordable rental units. It also administers 0 Section 8 housing vouchers.
According to HUD, Housing Authority of the County of Dona Ana is determined to be a Very Small public housing authority, meaning it manages between 1 - 49 public housing units. Also according to the Department of Housing and Urban Development, the housing authority is designated as No units, meaning it administers Section 8 vouchers.
Comparing the housing assistance distribution of Housing Authority of the County of Dona Ana between Public Housing Units (100%) and Section 8 Housing Vouchers (0%) to that of all housing authorities in New Mexico, Housing Authority of the County of Dona Ana has a larger proportion of public housing units than the average housing authority. The housing authority's proportion of Section 8 vouchers under management is smaller than the average housing authority in New Mexico.Source United States Department of Housing and Urban Development (hud.gov, 2014)

subset: commoncrawl


text:
Feldman M, et al.
Sleisenger & Fordtran's Gastrointestinal and Liver Disease. 8th ed. St. Louis, MO: Mosby; 2005.
Ferri F ed.
Ferri's Clinical Advisor 2011. 1st ed. Philadelphia, PA: Mosby Elsevier; 2010.
Firestein ED et al.
Kelley's Textbook of Rheumatology. 8th ed. Philadelphia, PA: Saunders; 2008.
Goldman L, Ausiello D, eds.
Cecil Textbook of Internal Medicine. 23rd ed. Philadelphia, PA: Saunders; 2008.
Habif TP.
Clinical Dermatology. 4th ed. St. Louis, MO: Mosby; 2004.
Harrison's Principles of Internal Medicine.
15th ed. McGraw Hill; 2001.
National Institute of Arthritis and Musculoskeletal and Skin Diseases website. Available at:
Noble J.
Textbook of Primary Care Medicine. 3rd ed.
St. Louis, MO: Mosby; 2000.
Rakel R.
Textbook of Family Medicine
2007. 7th ed. Philadelphia, PA: Saunders Elsevier; 2009.
Rakel RE, Bope ET.
Conn's Current Therapy. 60th ed. Philadelphia, PA: Saunders Elsevier; 2009.
Scleroderma Foundation website. Available at:
This content is reviewed regularly and is updated when new and relevant evidence is made available. This information is neither intended nor implied to be a substitute for professional medical advice. Always seek the advice of your physician or other qualified health provider prior to starting any new treatment or with questions regarding a medical condition.
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In the future, 511 will give you driving directions for any given start and end point.
For now, 511 provides an easy link to MapQuest, where you can enter your start and end location to find the best route for your trip.
Driving Directions
© Copyright 2009 LA SAFE. All rights reserved.


text:
- Welcome to the 2014/15 season
- Season Calendar
- New Productions 2014/2015
- New Productions 2015/2016
- Revivals 2014/2015
- Revivals 2015/2016
- Recitals 2014/2015
- Recitals 2015/2016
- Concerts 2014/2015
- Concerts 2015/2016
- NOW! Opera for you / Events for children and young people 2014/2015
NOW! Opera for You 2015/2016
events for children
and young people
- Special Events 2014/2015
- Special Events 2015/2016
- Cast Changes
- Ensemble, guest artists and the Opera Team
- The orchestra
- Opera Studio
- Videos
- Press Releases
Michael Porter
During Michael Porter's first year as a member of Oper Frankfurt's Opera Studio roles for the company included a Spirit / Sailor (Dido and Aeneas), Schmidt (Werther), Thibault (Die Sizilianische Vesper/The Sicilian Vespers) and 2nd Shepherd (Daphne). Roles during the 2014/15 season are the Vision of a Young Man (Die Frau ohne Schatten), Rodrigo (Otello), Brighella (Ariadne auf Naxos), Fourth Squire (Parsifal), Rudolf in a new production of Euryanthe and Sipho in the world premiere of Lior Navok's An unserem Fluss/By Our River. Before coming to Frankfurt Michael Porter was a member of the Opera Theatre in St. Louis' Young Artist Programme, where he sang a Mole / Juror (Alice in Wonderland), and the Seagle Music Colony's Young Artists' Programme, where roles included Benvolio (Roméo et Juliette) and Ferrando (Così fan tutte). He made his debut at Forth Worth Opera in 2013, singing Brighella in Ariadne auf Naxos. This American tenor studied at Indiana University.
Saison 2014/2015
(By Our River)
Lior Navok ARIADNE AUF NAXOS
Richard Strauss DIE FRAU OHNE SCHATTEN
Richard Strauss EURYANTHE
Carl Maria von Weber OTELLO
Giuseppe Verdi PARSIFAL
Richard Wagner


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This Christmas poem (my ninth) is the third of a trilogy of biblical "narrative" poems: the first being from Simeon's perspective and the second from Mary's. I determined last year that I would try to write a poem from the perspective of Jesus becoming incarnate -- becoming a man, and a baby -- yet remaining all the while God. It was quite a daunting challenge (writing as if one were God speaking!), but I tried to keep it relatively simple, for readers' sake and my own (those wishing to pursue theology proper and Christology -- the Incarnation and Two Natures of Christ -- more deeply, see my Holy Trinity web page). I hope you are edified and blessed by the poem and may God bless you and yours abundantly during this Advent / Christmas season.
Disclaimer for certain wary Protestants: if you don't like the "Mary / Catholic stuff"; just omit the second stanza in your reading. That will eliminate, I believe, any theological objection you may have, and the poem still works okay without it (as a five-stanza piece); but you'll be missing a crucial biblical and theological aspect of the Incarnation (including the Virgin Birth).
* * * * *
With My Father I created the universe: all things that are.
We decreed to save mankind by sacrifice of "God's Lamb";
Foretold in Isaiah 53: Messiah scorned, slain, and scarred.
My mother gave I the grace to be immaculate, without sin;
Fit vessel for God incarnate: ark of the new covenant pure.
Gabriel hailed New Eve "full of grace": redemption to begin;
Virgin with child by the Holy Spirit; man's salvation now sure.
In fullness of time I came down from glorious heaven above;
Clothed with human flesh: a baby-king (!) of Israel from birth.
Lying in a cave; a manger, in the world I had made and loved,
I was worshiped by shepherds and wise men filled with mirth.
Though seeing through infant eyes: helpless, innocent, meek;
I knew all things, possessed God's wisdom even in that hour.
As a baby, though King and Messiah, I cried and didn't speak;
But I sustained the entire universe by the Word of My power.
Even animals at My birth sensed the wonder prophesied of old;
Mary and Joseph were filled with thankful happiness and joy.
The three kings gave Me gifts: frankincense, myrrh, and gold;
Shepherds adored, fell before God: now present as a baby boy.
Mary held Me close and rejoiced, under the star's shiny beam;
I was dependent on her as a son, though I was God all the while.
I contemplated the reason I was born, and how I would redeem.
But now all was contentment and peace, so I rested and smiled.
Written on 12 December 2009.


text:
Hey everyone! Wanted to check in and give you an update on the California Walk A Thon.
If you haven't already registered, there is still plenty of time! Register online: http://www.preeclampsia.org/wat.asp or, contact me by email: email@example.com
One exciting new bit of news:
The Santa Barbara Police Department will be there to provide fingerprint/ID kit cards to all children present at the walk. The Officers will be there at 9:30am. So get there early to register (registration begins at 9:00am) and then you'll have plenty of time to get your fingertprint cards and check out the nearby carousel.
Looking forward to seeing all of you on the 20th - please don't hesitate to contact me with any questions!


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Updated: iPhone 3.0 Spotlight Search Finds Deleted Messages?
UPDATE: iPhone 3.1 does indeed fix this.
Matt Janssen has discovered what looks like a bug in iPhone 3.0 Spotlight, whereby the search feature keeps emails in the index/cache even after they've been deleted from the inbox and purged from the trash (and even obliterated from the host server). Janssen claims in the video above he's found emails he deleted several months ago using this method.
Email is likely cached to speed up searches. Initially tapping on a deleted email in a search result will launch Mobile Mail, which will promptly crash, but searching and tapping a second time will actually show the email, albeit in a buggy manner.
Engadget (via CultofMac) has heard this bug might be fixed in iPhone 3.1, now on Beta 3, and perhaps to be released as soon as Apple's next iPod/iTunes event rumored for the week of September 7.
Have something to say about this story? Leave a comment! Need help with something else? Ask in our forums!
Reader comments
Updated: iPhone 3.0 Spotlight Search Finds Deleted Messages?
Fastlane and Frag:
I remember the headlines on this a from WEEKS ago, I think they were even posted on the big geek sites (engadget, gizmodo) and they are running the same news ass if it were unheard of.
I guess nothing makes headlines like the iPhone!
This is a fantastic summary, I located your web page looking around aol for a related subject matter and arrived to this. I couldnt get to much other information on this piece of writing, so it was awesome to locate this one. I will end up being back again to check out some other posts that you have another time.


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Organizing Grid Set
Customers Also Bought
Organizing Grid Set Transforms Any Wall into Instant, Space-Saving Storage
- Create convenient storage on any wall
- Organizer Grid Set includes two wall grids, eight grid-to-wall mounting brackets (screws not included) and six 6" long Organizer Grid Hooks
Clean up clutter and make it easy to find items when you need them by organizing tools, gloves and more with these modular grids. Perfect for storing flower supports in the shed over the winter. You'll find dozens of uses around the house — store skis in the basement, hardware in the garage, craft items in the hobby room.
- Powder-coated, 1/8" diameter steel wire
- Grids are 27-1/2" L x 7/8" D x 11" H with 1-3/8" openings
- Hooks are 1-1/16" W x 7" D x 1-1/4" H with a 5 lb. weight capacity
- Grids sit 7/8" from wall; grid wire spacing is 1-3/8" square
- Brackets spaced for mounting on walls with studs 16" or 24" on center
- Mounting hardware not included; recommend wood screw at least 2" long if screwing into studs
- Gardener's Supply Exclusive
REVIEW SNAPSHOT®by PowerReviews
- Easy to install (4)
Best Uses
Reviewed by 9 customers
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Comments about Organizing Grid Set:
We have a plague of wasps here in SE Texas. I needed to organize my shed, but pegboard was not going to work, just more space for wasp's nests. This works perfect. Very strong, attached to the studs.
- Good For Organizing
- Sturdy
- Needs more hooks
Best Uses
Comments about Organizing Grid Set:
It definitely needs more hooks if you want it as an organizer.Would have ordered more hooks, if I knew.I use it as a craft and sewing organizer.Might not be a problem if you have other types of organizers to go with it.
- Easy To Install
- Holds a lot
- Like the look of it
- Needs more hooks
Best Uses
- Organize garage
Comments about Organizing Grid Set:
I used it organize my garage.
- Easy to intall
- Easy to use
- Sturdy
- Not for heavy items
Best Uses
- Garage
- Laundry Room
Comments about Organizing Grid Set:
I have these in the laundry room and garage and they are great for organizing lightweight items.
- Strong versatile
Best Uses
- Organizing small tools
Comments about Organizing Grid Set:
I am using it to organizing my small hand tools for gardening. The size of grids fits each of the tools and the pegs fits most of the tools. I have added carabiner rings to hold the tools securely.
- Easy To Install
Best Uses
Comments about Organizing Grid Set:
I put this up in my sewing room, with the shelves, to display large cones of thread.
I can hang extra scissors and rulers from the hooks and the baskets are great for CD/DVD storage.
- Easy To Install
- Easy to setup
- Saves space
- Very affordable
Best Uses
- Garden Tools
- Household items brooms
Comments about Organizing Grid Set:
I had tried the big box solutions for space savings but found them to be very expensive and difficult yo install.. THe organizing grid set solved all my problems!
Comments about Organizing Grid Set:
I wish it was eaier to mount on the wall.
- No Screws Included
- Parts come separate
- small
Best Uses
Comments about Organizing Grid Set:
I bought this for my dads man cave garage. I was disappointed when it arrived as it it pretty small, doesn't come with screws to attach to the wall, and kinda flimsy/cheap looking. Probably fine for very small, light items.
Product InstructionsInstructions for Organizing Grid Set
There are no special shipping restrictions or charges associated with this item. For detailed information about our delivery methods and charges, click here.
We're proud of the fact that this item is shipped to you directly from our warehouse in Vermont, by one of our 250 employee-owners. In-stock items are packed with care and shipped within the next business day to ensure prompt delivery to your home and garden!


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Tuesday, April 16, 2013
Three Boston Marathon runners from Summerville were not injured Monday in the explosion that shocked the nation.
The Summerville runners were Maureen Bodkin, Jessica Farmer and Lauren Jaynes. Official reports show that Jaynes finished in 3:42:26 and Bodkin in 4:01:04, putting her at the finish line just minutes before the 4:09:45 explosion.
Farmer is not shown as finishing the race, although she is shown as having passed her last checkpoint at 4:21:22.
Two explosions were reported. All runners who had not finished the race before the explosions were diverted off course.
According to Dorchester School District Two Public Information Officer Pat Raynor, Bodkin is employed at Beech Hill Elementary School as a school nurse.
Raynor said that Bodkin talked to a Beech Hill staff member Monday afternoon; the staff member said that Bodkin had finished the race and was about a block from the scene of the explosion with her son, who had accompanied her to Boston.
Bodkin and her son were not injured, and Bodkin is expected back at work at Beech Hill Wednesday, Raynor said.
A total of 130 South Carolinians took part in the race, according to reports.
Summerville resident Abby Caputo said that she and her mom stood across the street from the spot where the bomb went off when they attended the Boston Marathon two years ago to cheer on her dad who was running in the race.
Caputo has completed seven marathons but has never run the Boston Marathon.
"Running the Boston Marathon is a milestone for runners," Caputo said. "I remember standing there seeing the look of excitement and happiness on the faces of the runners, and the anticipation of those waiting for them at the finish line, and the thought of that being shattered by an event such as this is heartbreaking."
According to Caputo, runners qualify for the race through an elite process by running a qualifying time in what are known as "Boston qualifiers," and that if you are a runner it is something to which you aspire.
Chris May and Jacob Driggers of Charleston finished the race about 90 minutes before the explosions. They were back in their hotel when the blasts occurred.
Julia Early of Lexington had finished the marathon and was waiting to get a bag from a bus when she heard an explosion. Early says it sounded like a cannon. She says she made it back to her hotel room and was able to let family and friends know she was safe.
Amanda Perkins of Greenville says she had come across the finish line about four minutes before the first explosion. Perkins says she turned around and saw a gray cloud. She says there was no panic until the second blast.
Three people were confirmed dead in the blast, including an 8-year-old boy who was a spectator. Over 140 people were injured, and 17 were listed in critical condition.
The Associated Press contributed to this story.
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Wednesday, July 3, 2013
East Cooper Habitat held its 2013 House Party on May 17 at the Cooper River Room. A good time was had by all and many generous donations were received.
Moultrie News is pleased to offer readers the enhanced ability to comment on stories. We expect our readers to engage in lively, yet civil discourse. We do not edit user submitted statements and we cannot promise that readers will not occasionally find offensive or inaccurate comments posted in the comments area. Responsibility for the statements posted lies with the person submitting the comment, not Moultrie News.
If you find a comment that is objectionable, please click "report abuse" and we will review it for possible removal. Please be reminded, however, that in accordance with our Terms of Use and federal law, we are under no obligation to remove any third party comments posted on our website. Read our full terms and conditions.
© 2015 Moultrie News an Evening Post Industries company. All Rights Reserved.
Registration on or use of this site constitutes acceptance of our Terms of Service, Privacy Policy and Parental Consent Form.


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Wind: 0 mph
To The Editor:
I would like to echo and extend Dinsmore Fulton's points made in her letter last week (January 31, 2013). We are an energy consuming country and we are consuming more energy than ever, not less. We need to discuss rationally and honestly where we want that energy to come from.
I personally would like to see much more attention paid to reducing consumption, such as by increasing public support for public transportation, graduating the cost of commercial and residential energy usage as consumption increases and mandating greener building codes and much higher fuel efficiency for vehicles. However, I know reducing consumption alone will be insufficient since our usage continues to increase as we continually develop "smarter must have" technologies.
We need to confront this challenge by honestly discussing the pros and cons of all types of energy production and use. There are no perfectly clean and non-impactful energy options. We need to decide which ones strike the best short- and long-term balance between social, environmental and economic costs and benefits, in our community and nationally and internationally. But burying our heads in the sand or avoiding the problem with "not-in-my-backyard" answers that simply pass the buck to other communities are not appropriate responses.
Anne Bordonaro


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iFixit has done its usual trick of hopping over to Australia to get its hands on an iPad Air in the first time-zone to open its doors for business to bring us a look at the innards of the new device. The device is now on sale in the U.S. too, with supplies expected to be good.
No surprise that the company found little prospect of success for DIY repair, reporting that even opening the casing was a challenge: when you pack that much technology into so small a space, there's going to be a lot of glue involved.
Some details of what the company found and more photos below the fold …
The teardown confirmed Apple's battery-life magic: the company has indeed replaced the three-cell 43 WHr battery of the iPad 4 with a less-powerful two-cell battery (32.9 WHr) to reduce the dimensions, while the more efficient A7 and M7 chips allow even greater battery-life.
The A7 processor, though not given an X designation by Apple, is revealed to be a slightly different chip to the one on the iPhone 5S: marked as an APL5698 rather than APL0698.
The Broadcom touchscreen controllers are found to be similar to those used in MacBook trackpads, while last year's Micro-SIM is of course replaced by a Nano-SIM – all part of that quest to do everything possible to reduce the overall size of the tablet.
Repairability? 2 out of 10, mostly thanks to masses of glue and sticky-tape – again, the price you pay for the ultra-slim design.
Check out the full details over at the iFixit site. As an aside for European readers, iFixit now has a European store, offering faster and cheaper shipping on its repair kits.
Leave a Reply
16 Responses to "iPad Air teardown: never mind the repairability, feel the tech"
It will be interesting to see the GPU benchmarks on this. Hopefully Apple have used the top end 6 core PVR Rogue instead of the 4 core Rogue in the 5S. The iPad has a lot more pixels to fill, and will need the extra power.
Quite amusing to find out the iPad is held togther with glue and sticky tape
Why? Aircraft are, cars manufacturers use glue more and more… It has nothing to do with 'lesser quality'. Nowadays glue is high tech and the thing we design are so compact screws and bolts are no longer the right choice.
If you read a little more closely you'll see the poster was denoting humor…Humor, born it Britain but drowned trying to cross the channel…


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The Rosen Library will be be closed on Sunday, September 1st and Monday, September 2nd, in observance of Labor Day.
Normal Fall Hours will resume on Tuesday, September 3rd. For more information, visit the Rosen Library Hours page or Ask a Rosen Librarian.
Rosen Library Homepage | Ask a Rosen Librarian | Rosen Library Hours


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Local author pens Heisman biographyWritten by Sarah Ottney | Editor in Chief | firstname.lastname@example.com
At least once a week, John M. Heisman is asked if he is related to the legendary football coach.
"I'll go to a checkout, plop my card down and [the clerk] will say, 'Oh, Heisman. Like the trophy.' About 50 percent of the time, they'll say 'You're not related, are you?' Then I have a decision to make. Do I lie and say no or do I tell the truth and, if they are a sports fan, get into a time-wasting discussion? If I got back all the time I've spent answering that question, I'd have another year of my life."
The answer, of course, is yes.
John W. Heisman, for whom the Heisman Memorial Trophy was named, was the Toledo resident's great-uncle. The trophy is awarded each year to the nation's most outstanding college football player. Heisman has spent the past several decades researching and writing a book about his relative.
"Heisman: The Man Behind the Trophy" will be published Oct. 2 by Simon & Schuster. The book was co-authored by ESPN's Mark Schlabach with a foreword by University of South Carolina coach Steve Spurrier. A prerelease book-signing is set for 2-8 p.m. Oct. 1 at the Ward Pavilion at Wildwood Metropark, 4830 W. Central Ave.
"One of the things we tried to bring out in the book was the fact that his innovation created the American college football game as we know it now," Heisman said. "There were other contributors — we're not taking away from them — but he was a catalyst at a critical time in its development."
Spurrier, who won the Heisman Trophy in 1966, said he was surprised to learn just how much Heisman contributed to the sport.
"Among other innovations, Heisman introduced the center snap, audible 'hike' signal, hidden ball trick and lateral passes," Spurrier wrote. "Much of what Heisman invented nearly 100 years ago is still very much a part of the game today, which is quite remarkable."
The book was also endorsed by a dozen other Heisman winners, including former University of Michigan receiver Desmond Howard, who won the Heisman in 1991.
"As recognizable as the statue is, many do not know the storied history of the man it was named after," Howard wrote. "This book explores the life of a legend."
Coach Heisman played football for Brown University and the University of Pennsylvania, from which he graduated with a law degree.
"He was going to be an attorney, except for an injury to his eyes that kept him from reading for long periods, so he went to his next love — or actually his first love — which was athletics and football," Heisman said.
His first coaching job was at Oberlin College in 1892. Heisman also coached at Buchtel College in Akron (now the University of Akron), the Agricultural and Mechanical College of Alabama (now Auburn University), Clemson College (now Clemson University), Georgia Tech, which he led to a national championship in 1917, the University of Pennsylvania, Washington & Jefferson College and Rice Institute (now Rice University).
Grades and character were just as important to the coach as playing ability, Heisman said.
"If they used profanity, he pulled them out of the game and sent them to the showers. They were done," Heisman said. "He expected them to be gentlemen first, football players second."
Heisman never met the coach, who died before he was born, but remembers a visit from Heisman's wife, Edith. He grew up listening to his father talk about Heisman, whom the family called "Uncle Bill," after his middle name, William.
"When Dad wanted to give a lesson in character, I heard a story about Uncle Bill. When he wanted to impart a story of manhood, I heard a story about Uncle Bill. When he wanted me to understand good sportsmanship, scholastic effort or gentlemanly behavior, I heard an Uncle Bill story," Heisman wrote. "These stories were effective, entertaining and always hit the mark."
The coach died in 1936. When his wife died in the 1960s, the coach's personal effects were passed to Heisman's father. Heisman started going through the papers in the 1970s, but stopped to marry and raise a family before beginning work again in earnest in 2003.
"I went through his trunks and put it all together. It took decades," he said. "It's so very surreal [to be finished]. I don't know that my psyche has allowed me to acknowledge it yet. It feels like I'm still working. I haven't really slowed down to enjoy this yet."
Heisman played football for Genoa High School in the 1960s, where he said he was "outstandingly average," and then played rugby at Ohio State University, experiencing a game much like the college football his great-uncle knew.
"It gave me an appreciation for where the sport came from," Heisman said. "If you take away Heisman's contributions, you basically have an orphan-child blend of soccer and rugby … and basically without the fan base we have now. It's not the game we have now, not anywhere close."
Heisman said the most surprising thing he learned was how active his great-uncle was in theater — regularly acting in plays as well as managing an acting troupe during the offseason.
"For years he actually made more money on stage than he did coaching," Heisman said. "He was very shrewd and quite an entrepreneur. This guy went out, sized up opportunities and seized them. That was reflected in his coaching, in his acting and in how he went into communities and picked up sports columns and wrote for newspapers. He was sort of a Renaissance man."
Although Heisman doesn't usually attend the ceremony, he said he has become good at predicting the winner.
"I've been able to call it for the last several years, probably the last decade," he said. "There's only been a few surprises."
He's also met many of the winners.
"It would be easier to tell you the ones I haven't met," he said.
One of his most memorable interactions was with quarterback Tim Tebow, who won the Heisman in 2007.
"Tim looks you right in the eye. He's very genuine. He wants to know what you have to say," Heisman said.
Heisman was 12 years old the first time he saw a Heisman Trophy.
"It was pretty neat, but I've never been in awe of it because, if this is a memorial, then basically I'm looking at a tombstone," Heisman said. "That's the way I've always looked at it. It's too bad everyone has forgotten what the memorial is for and who they are honoring."
Heisman said he hopes the book sheds light on the history of football and the man behind the trophy.
"It's going to give a depth and a richness to the heritage and the tradition," John said. "I want people to understand a sense of history, a sense of the fact that this game we take for granted was basically a sandlot game back in the day."
Heisman also hopes the book will clear up his connection to the coach.
"Hopefully I won't be stopped at stores anymore and asked about my name," Heisman said, smiling. "That's my personal reason for writing this."
For more information, visit www.coachheisman.com.
Tags: Desmond Howard, Georgia Tech, Heisman Memorial Trophy, Heisman Trophy, Heisman: The Man Behind the Trophy, John M. Heisman, John W. Heisman, Mark Schlabach, Oberlin College, rugby, Sarah Ottney, Steve Spurrier, Tim Tebow


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Rapid Rewards at Southwest Airlines
Frances X. Frei; Corey Hajim
Price: $6.93
Used as part of the first module of a course on Managing Service Operations, which addresses managing the operating role of customers (606-032).
Southwest Airlines is well known as the low-fare airline that has achieved ongoing financial success in one of the most financially troubled industries in the United States. Told from the perspectives of two Southwest customers--a frequent flier and a more typical customer--the case revolves around two customer service requests from the frequent flier, both of which provide a compelling temptation to cater to the satisfaction of the airline's highest revenue customers. The requests are intriguing because they would neither add cost nor increase the turnaround time of Southwest's service.
Learning objective:
To explore the economics of the airline industry, specifically, the economic importance of frequent fliers; 2) To provide an opportunity to explore under what circumstances it makes sense for a company to differentiate its service offering across its customer base. Exploring the ramifications of these two customer service requests exposes students to the inherent tradeoffs and intricacies of Southwest's operating model and customer service philosophy.
Subjects Covered:
Customer service; Service management
- Industry: Airlines
- Company Employee Count:29,274
- Company Revenue:$5,650 million revenues
- Event Year Begin:2001
- Event Year End:2001
- Revision Date:
- Aug 23, 2004
- Publication Date:
- Aug 11, 2001
- Discipline:
- Service Management
- Source:
- Harvard Business School
- Product #:
- 24p, English PDF
- Also Available in:


text:
Ryan Scott Ottney
PDT Staff Writer
An Associated Press analysis of government data concluded that the number of people seeking treatment could double from current levels, depending on how many states expand their Medicaid programs and how many addicts pursue the new opportunity.
Ed Hughes, executive director of The Counseling Center in Portsmouth, called this "a good problem to have." He said Medicaid expansion in Ohio would extend coverage to 5,000 people in Scioto County, and The Counseling Center expects to pick up about 500 new drug treatment patients.
It has been 60 years since doctors concluded that addiction was a disease that could be treated, but today the condition still dwells on the fringes of the medical community. Only one cent of every health care dollar in the United States goes to addiction, and few alcoholics and drug addicts get care. One huge barrier, say many experts, has been a lack of health insurance. But that barrier crumbles in less than a year. In a major break with the past, 3 million to 5 million people with drug and alcohol problems — from homeless drug addicts to working mothers who drink too much — suddenly will become eligible for insurance coverage under the new health care overhaul.
Many with substance problems are waiting eagerly for January, when the new insurance will become available. The new demand could swamp the system before even half of the newly insured show up at the door, causing waiting lists of months or longer.
"It's the chance to clean up and not use anymore, so I could live a stable life," said Ashley Lore, 30, of Portsmouth told the Associated Press.
Lore was jailed and lost custody of her 4-year-old daughter as a result of her heroin addiction.
"If I get into treatment, I get visitation to my daughter back. And I get her back after I complete treatment," she said.
The AP found that those seeking a new chance at sobriety may be surprised by the reality behind the promise. The system for treating substance abuse — now largely publicly funded and run by counselors with limited medical training — is small and already full to overflowing in many places. In more than two-thirds of the states, clinics are already at or near capacity.
Only about 10 percent of the 23 million Americans with alcohol or drug problems receive treatment, according to the National Survey on Drug Use and Health. Shame and stigma are part of the reason, but about a quarter of them have no insurance. Today, those without insurance include many lower- and middle-income people who don't get the benefit from an employer, don't qualify for Medicaid or Medicare, and can't afford their own policies. The new law will provide subsidies to help many buy private coverage.
"It would be challenge, but it would be a challenge that we would love to try and meet," Hughes said. "We have struggled for the whole 30 years that I have worked in this field to provide services to people who are uninsured or under-insured, and the opportunity to have that number of people with access to treatment would be a real blessing for them and for our community. Because what we are seeing is, not only would they be able to access drug and alcohol treatment, but they could access other health care as well. Our community ranks 88 out of 88 counties in terms of poor health care indicators and our workforce is very sick and has drug addiction problems. If we are going to make progress on an economic front, we are going to have to have a healthier workforce."
The government is also pressing states to expand their Medicaid programs to include more working poor people. If 20 states expand Medicaid programs - roughly the number now planning to do so - 3.8 million more addicted patients would get insurance, the AP analysis found.
Gov. John Kasich supports Medicaid expansion in Ohio, but has been stymied by the Ohio Senate who have dropped the plan from the proposed state budget. Senate President Keith Faber said Wednesday that he's pushing for changes to the Medicaid program instead of the wide-ranging expansion plan.
Ryan Scott Ottney can be reached at 740-353-3101, ext. 287, or email@example.com. For breaking news, follow Ryan on Twitter @PDTWriter. The Associated Press contributed to this story.


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As the fifth season of The Apprentice starts tonight on NBC, Virender Ajmani is once again busy creating a new Google Maps mashup. This time he has created a Google "TV Map" mashup for the show's viewers to learn more about the candidates participating in this season's series and where they are from. Check it out!
With all these TV Maps popping up, I wonder how long it will take someone to create one to track The Amazing Race 9?? :)
Technorati Tags: [Apprentice], [Google Maps API], [Google], [Google Local], [Google Maps mashup], [Google Maps], [Google Maps Mania]


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A delegation from West Virginia came away from a meeting this month with the Environmental Protection Agency's new administrator feeling somewhat encouraged that they may be able to "hit the reset button" with federal regulators regarding the coal industry.
Whether that button is reset at all, or exactly in what place, remains to be seen. However, the group of Democrats — including members of the congressional delegation, Gov. Earl Ray Tomblin and others, as well as coal industry and labor representatives — said they believed new EPA chief Gina McCarthy at least heard their message.
If she truly did, that might be at least a beginning of a view in Washington that simply driving the coal industry into the ground has economic ramifications as well as an impact on the nation's energy picture.
Not long after Barack Obama took office as president in 2009, coal industry officials have maintained that his administration has waged a war on coal, with more stringent environmental regulations and holdups in the permitting of new mines. …
One is that the EPA's tough stance on coal does have an economic impact, contributing to a loss of jobs in the mining industry. It is not unreasonable to ask that EPA officials examine and consider that impact as they develop and enforce regulations.
Another is that coal will remain a key part of the nation's energy portfolio for many, many years to come, whether the powers-that-be in Washington like it or not. …
The meeting 10 days ago with McCarthy at least marked a step forward for West Virginia's relationship with the EPA. Her predecessor in that job wouldn't even acknowledge state leaders' letters asking for a meeting.
Now, it's a matter of waiting to see whether the dialogue with McCarthy will yield any policy changes in the EPA that result in a more balanced approach factoring in the economy as well as the environment.
— The Herald-Dispatch


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adventure time.
ok that's all.
Correct! Off work and heading for the hills avec le velo woo xReplyDelete
excellent georgie, love seeing those beautiful rollin green hills in your posts :)Delete
Makin pancakes, makin bacon pancakes.....ReplyDelete
OMG YES the best ever(!!) have u seen the alicia keys remix...Delete
gahaha love it!
.... can you share the alicia keys remix?
I read every comment, frequently with coffee in hand.
/ Sincere thanks for your readership. - - - - - - - -


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This large, modern, purpose-built hotel stands close to the town centre. It provides spacious, smartly presented and well-equipped bedrooms, all of which contain convertible bed settees for family occ...More about Novotel Wolverhampton
This delightful Cotswold country house has been appointed to provide the highest levels of quality, comfort and relaxation. Individually styled bedrooms come in a range of shapes and sizes, from the l...More about Barnsley House
This characterful inn has plenty of history and is located in the centre of the town, just one minute's walk from the high street. The Kings Arms is a traditional inn where access to some of the bedr...More about Kings Arms
High quality, budget accommodation ideal for both families and business travellers. Spacious, en suite bedrooms feature tea and coffee making facilities, and Freeview TV in most hotels. Internet acces...More about Premier Inn Kidderminster
High quality, budget accommodation ideal for both families and business travellers. Spacious, en suite bedrooms feature tea and coffee making facilities, and Freeview TV in most hotels. Internet acces...More about Premier Inn Birmingham City Centre New Street
Located in a small hamlet just west of Monmouth, close to the Forest of Dean and the Wye Valley, this operation offers accommodation comprising six very well-appointed cottages. The comfortable rooms ...More about The Stonemill & Steppes Farm Cottages
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An inn has occupied this site since the 13th century, though The Boot itself dates from the Georgian period. Much historic charm remains in the pub, while the bedrooms, furnished in antique pine, are ...More about The Boot Inn
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The present invention provides a method for rapid solution capture purification of nucleic acids for subsequent analysis by electrospray mass spectrometry which is efficient and cost-effective relative to existing methods. The present invention also provides for kits useful for practicing rapid solution capture of nucleic acids so that purified samples are in condition for analysis by electrospray mass spectrometry.
1. A method of purifying a solution comprising a nucleic acid comprising:
mixing said solution with an anion exchange resin to yield a suspension of said anion exchange resin in said solution wherein said nucleic acids bind to said anion exchange resin;
isolating said anion exchange resin from said solution;
washing said anion exchange resin to remove one or more contaminants with one or more wash buffers while retaining bound nucleic acids;
eluting said nucleic acids from said ion exchange resin with an elution buffer.
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26. A method of purifying a solution comprising a nucleic acid for subsequent analysis by electrospray mass spectrometry comprising:
adding a weak anion exchange resin to said solution and mixing to yield a suspension of said weak anion exchange resin in said solution wherein said nucleic acids bind to said weak anion exchange resin;
isolating said weak anion exchange resin from said solution;
washing said weak anion exchange resin to remove contaminants with one or more wash buffers while retaining bound nucleic acids; and
eluting said nucleic acids from said weak ion exchange resin with an elution buffer.
27. The method of
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30. A method of purifying a solution of nucleic acids for subsequent analysis by electrospray mass spectrometry comprising:
adding ProPac™ WAX weak anion exchange resin to said solution and mixing to yield a suspension of said weak anion exchange resin in said solution wherein said nucleic acids bind to said weak anion exchange resin;
isolating said weak anion exchange resin from said solution;
washing said weak anion exchange resin to remove contaminants with one or more wash buffers while retaining bound nucleic acids; and
eluting said nucleic acids from said weak ion exchange resin with an elution buffer comprising piperidine, imidazole and methanol.
31. The method of
32. A kit for purification of nucleic acids comprising:
a resin source plate containing anion exchange resin;
a sample mixing plate;
a filter plate or a microtube rack comprising a plurality of micro filter tubes;
one or more resin wash solutions; and
an ESI-MS-compatible elution buffer.
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This application is a continuation-in-part of U.S. application Ser. No. 10/844,938 filed May 12, 2004, and U.S. application Ser. No. 10/844,122 filed May 12, 2004, each of which claims priority to U.S. Provisional Application Ser. No. 60/470,547 filed May 13, 2003, each of which is incorporated herein by reference in its entirety.
This invention was made with United States Government support under DARPA/SPO contract BAA00-09 and under CDC grant 1 RO1 C1000099-01. The United States Government may have certain rights in the invention.
The present invention relates generally to the field of analysis of nucleic acids by mass spectrometry and provides methods and kits useful for this purpose.
Electrospray ionization mass spectrometry (ESI-MS) has become an important technique for the analysis of biopolymers. The multiple charging phenomenon allows fast, accurate and precise molecular mass measurement, identification of modifications and more detailed structural studies for very high-mass biopolymers.
Amplification of specific DNA sequences utilizing the polymerase chain reaction has widespread applications in many scientific disciplines, including microbiology, medical research, forensic analysis, and clinical diagnostics. Most often, PCR products are "sized" using traditional biochemical techniques such as standard gel electrophoresis using either intercalating dyes or fluorescently labeled primers. The Taqman™ assay, which is widely used in a number of PCR-based diagnostic kits, confirms the presence (or absence) of a specific PCR product but provides no direct readout on the size of the amplicon. So-called "real-time" PCR devices, which measure the laser-induced fluorescence of the PCR product during the amplification cycles, are used to quantify the amplification of DNA from a given DNA template and primer set. These methods have limited utility for relatively small amplicons (less than 150 base pairs), owing to the proportionately high fluorescence background, and do not provide any information with respect to amplicon heterogeneity or exact length.
Compared to these more traditional methods, mass spectrometry has several potential advantages as a platform on which to characterize PCR products, including speed, sensitivity, and mass accuracy. Because the exact mass of each of the bases which comprise DNA is known with great accuracy, a high-precision mass measurement obtained via mass spectrometry can be used to derive a base composition within the experimentally obtained mass measurement uncertainty (J. Aaserud, Z. Guan, D. P. Little and F. W. McLafferty, Int. J. Mass Spectrom. Ion Processes, 1997, 167/168, 705-712. and D. C. Muddiman, G. A. Anderson, S. A. Hofstadler and R. D. Smith, Anal. Chem. 1997, 69, 1543-1549). Methods for rapid identification of unknown bioagents using a combination of nucleic acid amplification and determination of base composition of informative amplicons by molecular mass analysis are disclosed and claimed in published U.S. Patent applications 20030027135, 20030082539, 20030124556, 20030175696, 20030175695, 20030175697, and 20030190605 and U.S. patent application Ser. Nos. 10/326,047, 10/660,997, 10/660,122 and 10/660,996, all of which are commonly owned and incorporated herein by reference in entirety.
Both MALDI (matrix assisted, laser desorption ionization) and electrospray (ESI) mass spectrometry have been employed to ionize PCR products for subsequent mass spectrometric detection. While MALDI is widely used to analyze short (20-mer or smaller) oligonucleotides, applications to amplicons in excess of 100 bp are less common. ESI is one of the most widely used ionization techniques for large biological molecules owing to the inherent "softness" of the ionization process, which allows DNA in excess of 500 bp to be ionized without dissociation.
In ESI, large charged droplets are produced in the process of "pneumatic nebulization" where the analyte solution is forced through a needle at the end of which is applied a potential sufficient to disperse the emerging solution into a very fine spray of charged droplets all of which have the same polarity. The solvent evaporates, shrinking the droplet size and increasing the charge concentration at the droplet's surface. Eventually, at the Rayleigh limit, Coulombic repulsion overcomes the droplet's surface tension and the droplet explodes. This "Coulombic explosion" forms a series of smaller, lower charged droplets. The process of shrinking followed by explosion is repeated until individually charged analyte ions are formed. The charges are statistically distributed amongst the analyte's available charge sites, leading to the possible formation of multiply charged ions conditions. Increasing the rate of solvent evaporation, by introducing a drying gas flow counter current to the sprayed ions, increases the extent of multiple-charging. Decreasing the capillary diameter and lowering the analyte solution flow rate i.e. in nanospray ionization, will create ions with higher m/z ratios (i.e. it is a softer ionization technique) than those produced by "conventional" ESI and are of much more use in the field of bioanalysis.
Unfortunately, ESI requires relatively clean samples and is notoriously intolerable of cationic salts, detergents, and many buffering agents commonly used in biochemical laboratories.
The buffer system commonly employed in the polymerase chain reaction includes electrospray incompatible reagents such as 50 mM KCl, 2 mM MgCl2, 10 mM Tris-HCl, and each of the four deoxynucleotide triphosphates (dNTPs) at 200:M. Even the presence of relatively low concentrations of metal cations (less than 100:M) can significantly reduce MS sensitivity for oligonucleotides as the signal for each molecular ion is spread out over multiple salt adducts. Thus, in addition to removing detergents and dNTPs, effective ESI-MS of PCR products requires that the salt concentration be reduced by more than a factor of 1000 prior to analysis.
Ethanol precipitation has been used to desalt PCR products for subsequent MS analysis as short oligonucleotides and salts are removed while the sample is concentrated (M. T. Krahmer, Y. A. Johnson, J. J. Walters, K. F. Fox, A. Fox and M. Nagpal, Electrospray Anal. Chem. 1999, 71, 2893-2900; T. Tsuneyoshi, K. Ishikawa, Y. Koga, Y. Naito, S. Baba, H. Terunuma, R. Arakawa and D. J. Prockop Rapid Commun. Mass Spectrom. 1997, 11, 719-722; and D. C. Muddiman, D. S. Wunschel, C. L. Liu, L. Pasatolic, K. F. Fox, A. Fox, G. A. Anderson and R. D. Smith Anal. Chem. 1996, 68, 3705-3712). In this method, the PCR product can be precipitated from concentrated ammonium acetate solutions, either overnight at 5� C. or over the course of 10-15 min with cold (−20� C.) ethanol. Unfortunately, a precipitation step alone is generally insufficient to obtain PCR products which are adequately desalted to obtain high-quality ESI spectra; consequently, precipitation is generally followed by a dialysis step to further desalt the sample (D. C. Muddiman, D. S. Wunschel, C. L. Liu, L. Pasatolic, K. F. Fox, A. Fox, G. A. Anderson and R. D. Smith Anal. Chem. 1996, 68, 3705-3712). While several researchers have successfully employed these methods to characterize a number of PCR products, the route to applying these methods in a robust and fully automated high-throughput manner is not obvious.
Commercial DNA purification kits may also be used in conjunction with traditional desalting techniques such as microdialysis (S. Hahner, A. Schneider, A. Ingendoh and J. Mosner Nucleic Acids Res. 2000, 28, e82/i-e82/viii; and A. P. Null, L. T. George and D. C. Muddiman J. Am. Soc. Mass Spectrom. 2002, 13, 338-344). Other purification techniques, such as gel electrophoresis followed by high-performance liquid chromatography or drop dialysis, or cation exchange using membranes or resins have also been used to obtain high-purity, desalted DNA for MS detection (L. M. Benson, S.-S. Juliane, P. D. Rodringues, T. Andy, L. J. Maher III and S. Naylor, In: The 47th ASMS Conference on Mass Spectrometry and Allied Topics, Dallas, Tex. (1999); C. G. Huber and M. R. Buchmeiser Anal. Chem. 1998, 70, 5288-5295; H. Oberacher, W. Parson, R. Muehlmann and C. G. Huber Anal. Chem. 2001, 73, 5109-5115; and C. J. Sciacchitano J. Liq. Chromatogr. Relat. Technol. 1996, 19, 2165-2178). Unfortunately, as with the techniques described above, the path toward a rapid and fully automated high-throughput implementation is not obvious.
Jiang and Hofstadler have developed and reported a single protocol for the purification and desalting of PCR products which employs commercially available pipette tips packed with anion exchange resin (Y. Jiang and S. A. Hofstadler Anal. Biochem. 2003, 316, 50-57). This protocol yields an ESI-MS-compatible sample and requires only 10:1 of crude PCR product. However, the method is cost-prohibitive when applied to high volume and high throughput processes such as the methods cited above for identification of unknown bioagents. Retail costs of using the commercially-obtained ZipTip™ AX (Millipore Corp. Bedford, Mass.) are estimated at $1.77 per plate well.
There remains a need for a method of purification of nucleic acids for mass spectrometry which is rapid, efficient and non-cost prohibitive. The present invention satisfies this need.
Solution capture of nucleic acids such as those obtained from amplification reactions has enabled a rapid, cost-effective method of extracting and purifying these analytes for subsequent analysis by mass spectrometry. Since the nucleic acids and the anion exchange media are in solution, efficient capture of the nucleic acids is accomplished by vortexing, or other mixing methods. This has eliminated the need to pack the media in a column format which would require multiple passes of the nucleic acid solution over it to achieve high levels of recovery of nucleic acids. While longer columns require fewer passes, significant backpressure becomes a problem. The process of packing an anion exchange resin in a column or pipette tip format increases the cost associated with the procedure accordingly. Thus the use of solution capture for purification of PCR products for analysis by mass spectrometry has substantially reduced the cost associated with sample preparation by eliminating the need to pack, equilibrate, and test a column. The retail cost of the current procedure using a pipette tip packed with anion exchange resin exemplified by ZipTip™ AX (Millipore, Bedford, Mass.) is approximately $1.77 per pipette tip (for each sample). The estimated cost of solution capture of PCR products is $0.10 per sample and takes into account the combination of anion exchange resin and filter plate. Furthermore, the time required for solution capture purification of PCR products is approximately 10 minutes per 96 well plate in contrast to the previous method which employs the ZipTip™ AX pipette tips and requires approximately 20 minutes.
The present invention is directed to, inter alia, solution capture methods of purifying a solution comprising one or more nucleic acids for subsequent analysis by electrospray mass spectrometry, or any other analysis, by adding an anion exchange resin to the solution and mixing to yield a suspension of the anion exchange resin in the solution wherein the nucleic acid binds to the anion exchange resin, isolating the anion exchange resin from the solution, washing the anion exchange resin to remove one or more contaminants with one or more wash buffers while retaining bound nucleic acid, eluting the nucleic acid, from the ion exchange resin with an elution buffer, and optionally, analyzing the nucleic acids by electrospray mass spectrometry.
The anion exchange resin may have a strong anion exchange functional group such as a quaternary amine or a weak anion exchange functional group such as, for example, polyethyleneimine, charged aromatic amine, diethylaminomethyl, or diethylaminoethyl. Such weak anion exchange resins comprise functional groups with pKa values of 9.0 or greater.
The present invention is further directed to kits for purification of nucleic acids comprising a filter plate comprising a plurality of wells or a tube rack comprising a plurality of tubes, an anion exchange resin, at least one anion exchange wash buffer and an ESI-MS-compatible elution buffer.
One embodiment of the method of solution capture purification of nucleic acids for analysis by mass spectrometry, for example, is outlined in
In some embodiments, a strong cation exchange functional group, such as a quaternary amine for example, is employed as the functional group of the anion exchange resin. Additional strong anion exchange functional groups are known to those skilled in the art.
In other embodiments, a weak anion exchange functional group is a suitable anion exchange functional group, such as polyethyleneimine, charged aromatic amine, diethylaminomethyl, or diethylaminoethyl, for example, are employed as the functional group of the anion exchange resin. Such functional groups have pKa values of 9.0 or greater. Commercial products of weak anion exchange resin include, but are not limited to; Baker PEI, Baker DEAM, Dionex ProPac™ WAX, Millipore PEI, Applied Biosystems Poros™ PI.
In some embodiments, the mixing of the anion exchange resin into the solution of nucleic acids is effected by repeated pipetting, vortexing, sonication, shaking, or any other method that results in suspension of the anion exchange resin in the solution containing the nucleic acids.
In some embodiments, dry anion exchange resin is added directly to the solution of nucleic acids or contained within a microtube or the well of a micro filter plate into which the solution of nucleic acids is added prior to mixing. In other embodiments, the anion exchange resin is pre-hydrated and added directly to the solution of nucleic acids or contained within a microtube or a well of a microfilter plate into which the solution of nucleic acids is added prior to mixing.
In some embodiments, the anion exchange resin which contains bound nucleic acids is isolated from the solution by filtration. Filtration can be effected, for example, using a filter plate in a 96- or 384-well format which enables high-throughput purification of multiple samples, or in any other container or plurality of containers equipped with a filter. Other well format plates can also be used. Membranes useful for filtration include but are not limited to those composed of the following materials: polytetrafluoroethylene (PTFE), polyvinyldifluoro (PVDF), polypropylene, polyethylene, glass fiber, polycarbonate and polysulfone. Filtering may by accomplished by vacuum, centrifugation, or positive pressure displacement with fluids or gases, or any other method that effects the isolation of the anion exchange resin from the solution. Methods of filtering are well known to those skilled in the art.
In some embodiments, the anion exchange resin comprises an anion exchange functional group which is linked to magnetic beads. Such an arrangement enables a simpler isolation step (110) by eliminating the need for centrifugation, vacuum or positive pressure displacement which would necessitate the removal of the plate or microtube tube from the liquid handler deck. Instead, a magnetic field can be activated to compress the magnetic bead resin so that liquid can be aspirated off by the liquid handler. Methods of using magnetic beads to effect isolation of biomolecules are well known to those skilled in the art.
In some embodiments, the anion exchange resin which contains bound nucleic acids is washed to remove one or more contaminants. Contaminants include, but are not limited to: proteins such as reverse transcriptase and restriction enzymes, polymers, salts, buffer additives, or any of the various components of an amplification reaction such as polymerases nucleotide triphosphates or any combination thereof. Depending on the composition of the contaminants in the nucleic acid solution, more than one wash buffer may be useful for removal of contaminants. Washing of the anion exchange resin can be effected with aqueous solutions of ammonium acetate in the millimolar range from about 20 mM to about 500 mM NH4OAc or with about 20 mM to about 500 mM NH4HCO3. Washing with about 10% to about 50% methanol, about 20% to about 50% methanol, or about 10% to about 30% methanol is useful as a final wash step. Methanol can be replaced by other suitable alcohols known to those skilled in the art.
In some embodiments, elution of nucleic acids from the anion exchange resin is accomplished using an ESI-compatible solution at alkaline pH of about pH 9 or greater such as an aqueous solution of about 2% to about 8% ammonium hydroxide or an aqueous solution of about 10 mM to about 50 mM, or 25 mM piperidine, about 10 mM to about 50 mM, or 25 mM imidazole and about 30% methanol or other suitable alcohol. As defined herein, an ESI-compatible solution is a solution which does not have a detrimental effect on the function of an electrospray (ESI) source.
As used herein, the term "about" means�10% of the term being modified. Thus, for example, "about" 10 mM means 9 to 11 mM.
In another embodiment, the present invention also provides kits for purification of nucleic acids by the solution capture method of the present invention. In some embodiments, the kit may comprise a sufficient quantity of anion exchange resin. In some embodiments, the anion exchange resin is a weak anion exchange resin such as one of the following commercially available weak anion exchange resins: Baker polyethyleneimine resin, Baker diethylaminomethyl resin, Dionex ProPac™ WAX, Millipore polyethyleneimine, and Applied Biosystems POROS™ PI.
In some embodiments, the kit may comprise a filter plate such as a 96- or 384-well filter plate or a microtube rack comprising a plurality of micro filter tubes.
In some embodiments, dry anion exchange resin is pre-loaded into the wells of a filter plate or microtube rack and can be either pre-hydrated or in the dry (powder) form.
The kit may also comprise a filter plate comprising a plurality of wells or a tube rack comprising a plurality of tubes, an anion exchange resin, at least one anion exchange wash buffer and an ESI-MS-compatible elution buffer.
In one embodiment, the kit may comprise a 96 or 384 well plate containing either pre-hydrated anion exchange resin or dry anion exchange resin, a second 96 or 384 well sample mixing plate, a 96 or 384 well filter plate, a resin treatment buffer, one or more wash buffers, and an ESI-compatible elution buffer.
In one embodiment, the nucleic acid solution is a PCR product prepared for identification of an unknown bioagent and contained in an individual well of a 96 well sample plate on the deck of an automated liquid handler. The liquid handler is the cornerstone for many laboratory processes associated with drug discovery and high throughput screening. The dispensing and aspiration functions of liquid handlers are used to perform solvent/reagent additions, dilutions, plate replications consolidation, redistribution and other microplate-based tasks and typically use disposable pipette tips for transferring liquids. Programming of liquid handlers to perform the various liquid handling tasks of this embodiment is well within the capabilities of one with ordinary skill in the art without undue experimentation.
The liquid handler is programmed to transfer and mix a predetermined volume of a suspension of anion exchange resin into the well containing the PCR product. The resin suspension can be contained in a resin source container such a 96 well plate and transferred to the PCR product plate by the liquid handler. Mixing is performed by the liquid handler via repeated dispensation and aspiration of the PCR-resin mixture and binding of nucleic acids to the resin occurs at this stage. Next, the liquid handler transfers the PCR product-resin mixture from the 96 well plate to a 96 or 384 well filter plate. At this stage, the filter plate can be removed from the liquid handler deck and the resin can be isolated from the solution by centrifugation or positive pressure displacement before returning the filter plate to the liquid handler deck.
The resin containing bound nucleic acids is then washed one or more times with an appropriate wash solution such as about 100 mM NH4HCO3 with the liquid handler pipetting the wash solution into the filter plate, followed by centrifugation, vacuum, or positive pressure displacement followed by one or more washes with about 20% to about 50% methanol before returning the filter plate containing the resin and bound nucleic acids to the liquid handler deck.
Finally, the nucleic acids are eluted from the resin with an ESI compatible elution buffer such as an aqueous solution of about 25 mM piperidine, about 25 mM imidazole and about 50% methanol. This ESI compatible buffer may also optionally contain an internal standard used to calibrate the ESI mass spectrometer during the subsequent ESI mass spectrometry analysis.
In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner. Throughout these examples, molecular cloning reactions and other standard recombinant DNA techniques were carried out according to methods described in Maniatis et al., Molecular Cloning—A Laboratory Manual, 2nd ed. Cold Spring Harbor Press (1989), using commercially available reagents except where otherwise noted.
In one embodiment, nucleic acid is isolated from the organisms and amplified by PCR using standard methods prior to BCS determination by mass spectrometry. Nucleic acid is isolated, for example, by detergent lysis of bacterial cells, centrifugation and ethanol precipitation. Nucleic acid isolation methods are described in, for example, Current Protocols in Molecular Biology (Ausubel et al.) and Molecular Cloning; A Laboratory Manual (Sambrook et al.). The nucleic acid is then amplified using standard methodology, such as PCR, with primers which bind to conserved regions of the nucleic acid which contain an intervening variable sequence as described below.
General Genomic DNA Sample Prep Protocol: Raw samples are filtered using Supor-200 0.2 μM membrane syringe filters (VWR International). Samples are transferred to 1.5 ml eppendorf tubes pre-filled with 0.45 g of 0.7 mm Zirconia beads followed by the addition of 350 μl of ATL buffer (Qiagen, Valencia, Calif.). The samples are subjected to bead beating for 10 minutes at a frequency of 19 l/s in a Retsch Vibration Mill (Retsch). After centrifugation, samples are transferred to an S-block plate (Qiagen) and DNA isolation is completed with a BioRobot 8000 nucleic acid isolation robot (Qiagen).
Swab Sample Protocol: Allegiance S/P brand culture swabs and collection/transport system are used to collect samples. After drying, swabs are placed in 17�100 mm culture tubes (VWR International) and the genomic nucleic acid isolation is carried out automatically with a Qiagen Mdx robot and the Qiagen QIAamp DNA Blood BioRobot Mdx genomic preparation kit (Qiagen, Valencia, Calif.).
The mass spectrometer used is a Bruker Daltonics (Billerica, Mass.) Apex II 70e electrospray ionization Fourier transform ion cyclotron resonance mass spectrometer (ESI-FTICR-MS) that employs an actively shielded 7 Tesla superconducting magnet. All aspects of pulse sequence control and data acquisition were performed on a 1.1 GHz Pentium II data station running Bruker's Xmass software. 20 μL sample aliquots were extracted directly from 96-well microtiter plates using a CTC HTS PAL autosampler (LEAP Technologies, Carrboro, N.C.) triggered by the data station. Samples were injected directly into the ESI source at a flow rate of 75 μL/hr. Ions were formed via electrospray ionization in a modified Analytica (Branford, Conn.) source employing an off axis, grounded electrospray probe positioned ca. 1.5 cm from the metalized terminus of a glass desolvation capillary. The atmospheric pressure end of the glass capillary is biased at 6000 V relative to the ESI needle during data acquisition. A counter-current flow of dry N2/O2 was employed to assist in the desolvation process. Ions were accumulated in an external ion reservoir comprised of an rf-only hexapole, a skimmer cone, and an auxiliary gate electrode, prior to injection into the trapped ion cell where they were mass analyzed.
Spectral acquisition was performed in the continuous duty cycle mode whereby ions were accumulated in the hexapole ion reservoir simultaneously with ion detection in the trapped ion cell. Following a 1.2 ms transfer event, in which ions were transferred to the trapped ion cell, the ions were subjected to a 1.6 ms chirp excitation corresponding to 8000-500 m/z. Data was acquired over an m/z range of 500-5000 (1M data points over a 225K Hz bandwidth). Each spectrum was the result of co-adding 32 transients. Transients were zero-filled once prior to the magnitude mode Fourier transform and post calibration using the internal mass standard. The ICR-2LS software package (G. A. Anderson, J. E. Bruce (Pacific Northwest National Laboratory, Richland, Wash., 1995) was used to deconvolute the mass spectra and calculate the mass of the monoisotopic species using an "averaging" fitting routine (M. W. Senko, S. C. Beu, F. W. McLafferty, J. Am. Soc. Mass Spectrom. 1995, 6, 229) modified for DNA. Using this approach, monoisotopic molecular weights were calculated.
For pre-treatment of ZipTips™ AX (Millipore Corp. Bedford, Mass.), the following steps were programmed to be performed by an Evolution™ P3 liquid handler (Perkin Elmer) with fluids being drawn from stock solutions in individual wells of a 96-well plate (Marshall Bioscience): loading of a rack of ZipTips™ AX; washing of ZipTips™ AX with 15 μl of 10% NH4OH/50% methanol; washing of ZipTips™ AX with 15 μl of water 8 times; washing of ZipTips™ AX with 15 μl of 100 mM NH4OAc.
For purification of a PCR mixture, 20 μl of crude PCR product was transferred to individual wells of a MJ Research plate using a BioHit™ multichannel pipette. Individual wells of a 96-well plate were filled with 300 μl of 40 mM NH4HCO3. Individual wells of a 96-well plate were filled with 300 μl of 20% methanol. An MJ research plate was filled with 10 μl of 4% NH4OH. Two reservoirs were filled with deionized water. All plates and reservoirs were placed on the deck of the Evolution™ P3 (EP3) pipetting station in pre-arranged order. The following steps were programmed to be performed by an Evolution™ P3 pipetting station: aspiration of 20 μl of air into the EP3 P50 head; loading of a pre-treated rack of ZipTips™ AX into the EP3 P50 head; dispensation of the 20 μl NH4HCO3 from the ZipTips™ AX; loading of the PCR product into the ZipTips™ AX by aspiration/dispensation of the PCR solution 18 times; washing of the ZipTips™ AX containing bound nucleic acids with 15 μl of 40 mM NH4HCO3 8 times; washing of the ZipTips™ AX containing bound nucleic acids with 15 μl of 20% methanol 24 times; elution of the purified nucleic acids from the ZipTips™ AX by aspiration/dispensation with 15 μl of 4% NH4OH 18 times. For final preparation for analysis by ESI-MS, each sample was diluted 1:1 by volume with 70% methanol containing 50 mM piperidine and 50 mM imidazole.
For pre-treatment of ProPac™ WAX weak anion exchange resin, the following steps were performed in bulk: sequential washing three times (10:1 volume ratio of buffer to resin) with each of the following solutions: (1) 1.0 M formic acid/50% methanol (2) 20% methanol (3) 10% NH4OH (4) 20% methanol (5) 40 mM NH4HCO3 (6) 100 mM NH4OAc. The resin is stored in 20 mM NH4OAc/50% methanol at 4� C.
Corning 384-well glass fiber filter plates were pre-treated with two rinses of 250 μl NH4OH and two rinses of 100 μl NH4HCO3.
For binding of the PCR product nucleic acids to the resin, the following steps were programmed to be performed by the Evolution™ P3 liquid handler: addition of 0.05 to 10 μl of pre-treated ProPac™ WAX weak anion exchange resin (30 μl of a 1:60 dilution) to a 50 μl PCR reaction mixture (80 μl total volume) in a 96-well plate; mixing of the solution by aspiration/dispensation for 2.5 minutes; and transfer of the solution to a pre-treated Corning 384-well glass fiber filter plate. This step was followed by centrifugation to remove liquid from the resin and is performed manually, or under the control of a robotic arm.
The resin containing nucleic acids was then washed by rinsing three times with 200 μl of 100 mM NH4OAc, 200 μl of 40 mM NH4HCO3 with removal of buffer by centrifugation for about 15 seconds followed by rinsing three times with 20% methanol for about 15 seconds. The final rinse was followed by an extended centrifugation step (1-2 minutes).
Elution of the nucleic acids from the resin was accomplished by addition of 40 μl elution/electrospray buffer (25 mM piperidine/25 mM imidazole/35% methanol and 50 nM of an internal standard oligonucleotide for calibration of mass spectrometry signals) followed by elution from the 384-well filter plate into a 384-well catch plate by centrifugation. The eluted nucleic acids in this condition were amenable to analysis by ESI-MS (See
To investigate the efficacy of the solution capture method of the present invention, the ESI-MS analysis results obtained for PCR products purified with the solution capture method (Example 4) were compared with the ZipTips™ method outlined in Example 3.
Bacillus anthracis DNA was isolated and amplified by PCR using a primer pair that amplifies a section of the lef gene of B. anthracis ranging from residues 756-872. Shown in
To confirm the efficacy of carrying out solution capture of nucleic acids with ion exchange resin linked to magnetic beads, 25 μl of a 2.5 mg/mL suspension of BioClon amine terminated supraparamagnetic beads were added to 25 to 50 μl of a PCR reaction containing approximately 10 pM of a typical PCR amplicon such as an amplicon obtained from broad priming of Staphylococcus aureus. The above suspension was mixed for approximately 5 minutes by vortexing or pipetting, after which the liquid was removed after using a magnetic separator to separate out the beads. The beads containing bound PCR amplicon were then washed 3� with 50 mM ammonium bicarbonate/50% MeOH or 100 mM ammonium bicarbonate/50% MeOH, followed by three more washes with 50% MeOH. The bound PCR amplicon was eluted with 25 mM piperidine, 25 mM imidazole, 35% MeOH, plus peptide calibration standards.
The eluate was then analyzed by ESI-FTICR electrospray ionization mass spectrometry (ESI). The ESI-FTICR mass spectrum is shown in
Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference (including, but not limited to, journal articles, U.S. and non-U.S. patents, patent application publications, international patent application publications, gene bank accession numbers, and the like) cited in the present application is incorporated herein by reference in its entirety.
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Mobile Nations Passport Contest: Win the Ultimate iOS Prize Pack!
Login to iMore and leave a comment to this post to be entered to win the Ultimate iOS Prize Pack!
Last week we rolled our new Mobile Nations Passport account system, which makes it easy for any member of iMore to also participate across our other sites. With Passport, you have one account and to rule them all!
In honor of the successful roll out of Passport and thanks to our awesome friends at Gogo, we're celebrating with an amazing cross site contest. Each of our sites is running a contest giving away an Ultimate Prize Pack for their platform. And thanks to Passport, you can easily enter for a shot at winning each prize pack on each site.
How to Enter:
Step 1: Login to iMore and leave a comment to this post to be entered to win the Ultimate iOS Prize Pack!
Register or Login using the links at the top right corner of this site (see where it says Passport Is Live)
Step 2: Also enter to Win the Ultimate Android, BlackBerry, Windows Phone and webOS Wildcard Prize Packs:
Click on over to Android Central, CrackBerry, Windows Phone Central and webOS Nation (<-- on those links) to go straight to the respective contest post for that site. Simply login on each site -- which is painless thanks to Passport -- and leave a comment there, as you're doing here, and you'll be entered to win that prize pack.
Step 3: Like Gogo on Facebook!
Our friends at Gogo -- the KINGS of Inflight Internet -- made it possible for us to run this Mobile Nations Contest. Gogo ROCKs. So be sure to click the button below and like them on Facebook. Your comment to this post is enough to be entered to win, but we'll love you, and Gogo will love you, if you like them (if the button doesn't work in your region, don't panic).
Beyond the Grand Prize, we have awesome runner up prizes as well. Keep reading for more info on the prizes and for the full contest details. Contest is open worldwide and ends at Noon PT on Monday, November 19th. Don't waste time... Enter now! Good Luck!
Mobile Nations Passport Contest - Full Contest Details
Prize Packs:
Click the Prize Pack links to jump over to that site to enter to win it!
Runner Up:
- Nexus 7 Tablet (32GB) - 5 Gogo passes
Runner Up:
- BlackBerry PlayBook (64GB)
- 5 Gogo passes
Runner Up:
- iPod Touch (32GB)
- 5 Gogo passes
Runner Up:
- JBL PowerUp Speaker
- 5 Gogo passes
Runner Up:
- Take your pick of any of the Runner Up Prizes we're offering for Android, iOS, Windows Phone and BlackBerry
Contest Rules & Fine Print:
Nothing to hide here, but these are the things to know!
- Contest is being run by Mobile Nations
- Only enter one comment per contest post on each site. If you enter additional comments, only your first comment will be counted towards entry of the contest.
- Contest is open worldwide.
- Contest ends at 12 Noon PT on Monday, November 19th, 2012.
- Contest winners will be drawn and random and announced on all Mobile Nations sites soon after.
- We will do our best to reach out to all contest winners. If a contest winner for whatever reason does not reply within a month of the winners announcement, Mobile Nations reserves the right to select a new winner for that contest prize.
- Individual prize winners are responsible for any taxes due on prize total if required by local laws.
- If the prize is not available at the time of winners announcement (example - a BlackBerry 10 phone), the prize will be delivered soon after it becomes available on the market for purchase.
Any questions? Email email@example.com.
Have something to say about this story? Leave a comment! Need help with something else? Ask in our forums!
Reader comments
Mobile Nations Passport Contest: Win the Ultimate iOS Prize Pack!
920 for the Win!
count me in!
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I would love some free stuff.... thanks
A free Apple product is the only good one :P
pick me
Fingers crossed! You never know, I suppose! :D
Woot! Nice work Mobile Nations.
Yes, even a WP fan likes an iPad and an iPhone, thanks very much!
this is amazing!
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very cool :-)
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Go go Gadget contest
ill take one
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You guys have the best contests
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Great contest - count me in!
I'd love to win!
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I would like to win this.
Did they announce the winners of that week long iphone 5 contest?
Winner Winner
I can't wait to win this and I think it is a great way to get me to look at the other mobile nations sites. Really cool and enlightening.
I Hope I am Selected
Yes, please!
Awesome! This is a sweet prize.Thanks for the chance!
This time , I hope I win one of these amazing prizes..
The chances of winning this are slim, but something is better than nothing!
the ultimate of all Ultimate Prize Packs is this one right here! Let's go!!
I would love to win this!
If i win Im going to give my mom a big hug
2 delicious prizes, and thousands of people to overcome :D here we go
wow awesome, contest!
Gotta say that when it comes to what's going on in the world of iOS there isn't a better place to visit. With my job I have to constantly be on top of what's new with iOS and this website has been a godsend. Thanks for keeping me in the loop and for all the hard work you guys do. All of the Mobile Nations websites are absolutely critical for me to succeed at what I do. And your contests aren't too shabby either. :)
I wouldn't mind having one of these!!
Great present for the wife
I'd love a free iPhone 5!
Mind boggling!! Count me in!
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Coming my way baby!
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iHope iWin.
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I hope I can win this time :/
That's a lot of awesome prizes!! Good luck to everyone. :) Mobile Nations is the best!!
iWin :D
OMG! What awesome prizes! Pick me! Please!!!!!
me please
This is the winning comment. Thanks for choosing me!
An unlocked iPhone 5... Hmm... Should I throw my sim card into it or use it to buy parts for my PC? Either way, I WANT TO WIN!!!!!
I would love to win this.
yay for free stufff =D
All digits crossed! Thanks for the amazing contest :D
sign me up!
iWant!! iWant!! iWant!! iWant!!
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hope I win
Great contest and great prizes!
I´ve posted on every mobilenation looking for a chance to win, s so if Rene choose my comment as one of the best, it would make me the happiest adolescent in the world =) & thanks to gogo for making this possible.I really want an iPhone 5, (I´ll also be happy if I win an iPod touch)
here Gabriel from Ecuador (the coutry conformed by the Galapagos Islands) hahaha #justincase
This is superb, thanks Mobile Nations!
I so so so want to win this. Fingers crossed, toes crossed, everything crossed. I never win anything, but glad that iMore is offering such great giveaways. Pleaseeeee I so want to winnnn the iOS Ultimate Prize Pack!!!!!!
I hope I win!! Thanks!!
Greatest prize ever!
Woot woot this is an awesome give away!
Mobile Nations!!!! Mobile Nations!!!!
Winner winner chicken dinner
Mmmm... free apple products.


text:
Roller Brushes
The roller brush of your vacuum cleaner sweeps dirt and debris off your floors. The bristles of the brush can be worn down over the period of time. This will prevent the brush from collecting all the dust and grime from the carpets and bare floors. The roller brush can be replaced if they are damaged or the bristles are worn out.
How to increase the life span of a roller brush
•The longetivity of the vacuum brush depends on how well you take care of it. The vacuum must be dusted and cleaned once in a while.
•Unfasten the faceplate over the brush bar and remove the plastic faceplate. Using a pair of scissors cut the threads and hair that are entangled around the brush bar.
•Using a rag wipe the dust and debris from the brush bar.
•Replace the faceplate and tighten the fasteners before using the vacuum.
Roller brushes of all the leading brands are available at GoVacuum.com. This is a place which offers replacements and the equipments at a bargain price. We also offer a world class customer service. Visit our site for all your unique cleaning needs. You can also chat with our customer care personal to clarify any of your queries.
Filter Queen Power Nozzle Brushroll 112C, AT1100$59.89Works with: Filter Queen 112C, Filter Queen AT1100 Learn More
Results 1 - 1 of 1


text:
Glendale mayor: No extension to be offered in Phoenix Coyotes deal
The clock is ticking on the potential deal between the City of Glendale and prospective Phoenix Coyotes owner Greg Jamison.
Less than 72 hours before the deadline for Jamison to buy the team from the National Hockey League, Glendale Mayor Jerry Weiers said there will be no extension for the proposed 20-year deal that would pay Jamison to manage the city-owned Jobing.com Arena.
Weiers wouldn't speculate on where the matter stands but said the city gave Jamison until Thursday and not "one second longer."
"We are not there yet," he said. "If he doesn't put the deal together, we won't go down that road again with him. Obviously, if we can keep the Coyotes here under different circumstances and without that same deal it is certainly something I would like to push forward with."
However, Arizona Sports 620's Paul Calvisi said the Coyotes deal is expected to be completed by Wednesday.
Update on #Coyotes sale to Greg Jamison:Multiple sources say "deal should be done by tomorrow" ...a day ahead of the deadline.— Paul Calvisi (@PaulCalvisi) January 29, 2013
The deal guarantees Jamison about $300 million over the next 20 years, should the purchase of the team be completed.


text:
The 11th Annual HORAN All For One Golf Classic is Monday, August 18
June 3, 2014
CINCINNATI - The 11th Annual HORAN All For One Golf Classic is set to take place on Monday, August 18. This year's event will feature a morning and afternoon golf flight at Kenwood Country Club and an afternoon flight at Hyde Park Country Club.
This annual outing serves as the signature fundraising event for the Xavier Athletic Department and helps to support all of Xavier's 16 varsity sports programs. Since its inception, the All For One Golf Classic has over $1,000,000 in net proceeds for Xavier Athletics.
Morning flight foursomes are $1,300.00 and afternoon flight foursomes are $1,600.00. The day includes golf & cart fees, food & drinks, golf gift package and all course games. Individuals will also receive one complimentary admission to the Night of Excellence event (August 16), which will serve as the official AFO Golf Classic dinner this year.
The AFO Night of Excellence event will take place on Saturday, August 16 (6 p.m.) at Cintas Center. The event will feature dinner, silent and live auctions and several exciting guest speakers that include Xavier Hall of Fame member and former Men's Basketball Coach, Pete Gillen. The program will also include a preview of the 2014-15 Men's and Women's basketball seasons by head coaches Chris Mack and Brian Neal. Like the HORAN All For One Golf Classic, all proceeds from this event benefit Xavier student-athletes and all of Xavier's varsity sports programs.
For more information, please visit www.xavier.edu/afogolf and if you have additional questions, please contact Tom Brady at (513) 745-4256 or email@example.com or Darlene Gilligan at (513) 745-3192 or email@example.com.


text:
Slider Revolution is an all-purpose slide displaying solution that allows for showing almost any kind of content with highly customizable, transitions, effects and custom animations. Due to its visual oriented interface and countless options, Slider Revolution is suited for beginners and pro's alike!
Slider Revolution is currently the top-selling slider plugin on Codecanyon and widely used by theme authors on Themeforest. Our team at ThemePunch is working hard on a daily basis to make this product better, faster and more usable. Updates are released on a regular basis and are of course totally free!
The latest version introduces many new modern slide transition types, including zoom and parallax types. Fancy 3D transitions are also available. If you prefer a more subtle approach, you can always select classical fade and slide transitions. There is definately something that fits your taste!
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A brand new feature of Slider Revolution is the ability to pull slider content from WordPress Posts and Custom Posts. You can create templates that determine how the title, date, etc. of a post is being displayed in form of captions using a flexible wildcard system. If you aren't into customizing, just select one of our premade post templates!
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Enjoy our highly improved Ken Burns effect in these sliders. Just import the demo sliders, add your own images and texts and you're done!
Slider Revolution cannot only handle Youtube & Vimeo Videos perfectly but also your self-hosted HTML5 Video! Check it out!
Another great example of whats possible wih Revolution Slider. Create unique Sliders with great usability. Try navigating this one with your arrow keys!
Here are some unique slider examples that show off what else is possible with the Slider Revolution plugin!
The possibilities of Slider Revolution are almost limitless with new features being added regularly. Here is a list of things we have included so far!
What do the experts think of Slider Revolution? Find out here!
Read a detailed review about Slider Revolution over at WPMU.org: http://wpmu.org/revolution-slider-a-premium-drag-and-drop-slider-for-wordpress/
Read about the "8 Must Have WordPress Plugins for Nonprofits" over at orgspring.com: http://orgspring.com/8-must-have-wordpress-plugins-for-nonprofits/
Read the "Revolution Slider WordPress Plugin Review & How To Guide" over at wpexplorer.com: http://www.wpexplorer.com/revolution-slider-review-guide/
Full Review of "Slider Revolution" at WP MAYOR: http://www.wpmayor.com/plugin-reviews/revolution-slider-review/
Wondering which license is best for you? Here is all the info you need!
Our most popular license, the Regular License allows use of the item in one single end product which end users are not charged to access or use. You can do this directly or, if you're a freelancer, you can create the end product for one client to distribute free to its end users. You can charge your client to produce the single end product. Distribution of source files is not permitted.
Our Extended License allows use of the item in one single end product, which you or one client may sell to end users (charge end users to access or use the end product). Distribution of source files is not permitted.
You can use Slider Revolution in your ThemeForest item if you purchase one extended license for that purpose. Meaning 1 Extended License for every single ThemeForest item you upload (1 ThemeForest Theme/Template Upload = 1 Extended License, 2 ThemeForest Themes/Templates Upload = 2 Extended Licenses etc.).
No. The extended license is still limited to a single end product, but you can re-sell that product.
Use of our items in stock projects is not allowed under the default regular or extended licenses. Contact us via our Profile page for further Permission and Discussion.