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Cicerone Guest House, Florence (province)
- Free continental breakfast and WiFi
Main amenities
- 8 guestrooms
- Room service
- Air conditioning
- Multilingual staff
- Luggage storage
- Tour/ticket assistance
What's around
- In San Lorenzo
- Cathedral of Santa Maria del Fiore (0.5 mi / 0.8 km)
- Academy Gallery (0.3 mi / 0.5 km)
I stayed here for one night. It was very clean and cozy. Also it was reasonable price.
Cicerone Guest House is a fantastically located 8-bedroom hotel. It takes about 5-8 mins…
Enter dates to check prices
Hotel Info: 020 3027 9803from £57
- Double Room Single Use
- Standard Triple Room
- Standard Double or Twin Room
- Family Room
Recommended hotels
At a glance
Key facts
Hotel size
- This hotel has 8 rooms
- This hotel is arranged over 1 floor
- Check-in time 2:00 PM-6:00 PM
- Check-out time is 11:00 AM
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
Travelling with others
- One child (18 years old and younger) stays free when occupying the parent or guardian's room, using existing bedding
Extra people
- Rollaway beds available, charges may apply*
- Free WiFi in public areas
- Free WiFi in rooms
Nearby One Star Florence (province) hotels
In the hotel
- Free continental breakfast daily
- Room service
- Tours/ticket assistance
- Luggage storage
In the room
- Climate control
- Air conditioning
- Desk
- Free WiFi
Top Florence (province) hotels
Recently reviewed hotels in Florence (province)
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"Thoroughly recommend the hotel, area and a few days in Florence!"Jun 22, 2015
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Cicerone Guest House, Florence (province)'s small print
Cash transactions at this property cannot exceed EUR 999.99, due to national regulations. For further details, please contact the property using information in the booking confirmation.
Mandatory fees
You'll be asked to pay the following charges at check in or check out:
- A tax is imposed by the city: EUR 2.50 per person, per night, up to 7 nights. This tax does not apply to children under 12 years of age.
Optional extras
Parking is available nearby and costs EUR 20 per 24 hours
Rollaway beds are available for EUR 30 per night
We have included all charges provided to us by this hotel. However, charges can vary, for example, based on length of stay or the room you book.


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Stock Photo:
young businessman on yellow wheat field
- Image ID: 66171415
- Copyright: Iakov Kalinin
- Release Information: Signed model release filed with Shutterstock, Inc
High quality images curated from over 150 countries.
Over 50 million photos, vectors, illustrations, and more.
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subset: commoncrawl


text:
New Faculty Welcome!
Human Resources is sponsoring the annual New Faculty Welcome! on September 18, from 9:15 a.m. to 4:00 p.m. in the Student Union Recreation Center (SURC) Ballroom.
New faculty will:
Click here to view the agenda and click on the campus map to locate the SURC building. Faculty, contact Jordyn Ashford, 509-963-2270, email@example.com, to confirm your attendance or to receive further information.
See the 2015 Employee Appreciation Awards Recipients!HR Newsletter - June/July Edition
Latest Edition of HR CentralJune/July 2015


text:
|Grant's Farm
10501 Gravois Rd
St. Louis, Missouri
Dining St. Louis,
a Fine Restaurant Guide
St. Louis Front Page
3832 S Broadway Street
St. Louis, MO 63118
To submit news, contact:
To advertise, contact:
ST. LOUIS, MO, (SLFP.com) - Grant's Farm has opened for its 61st season of family fun with traditional family favorites as well as new activities including:
"For more than 60 years Grant's Farm has provided St. Louisans with a unique, family-friendly atmosphere, making it a beloved local attraction," said Andy Elmore, general manager of Grant's Farm. "This year we're excited to welcome back visitors for another great season of fun, helping generations of guests make memories that last a lifetime."
Grant's Farm favorites like tram rides, baby goat feedings, behind-the-scenes Budweiser Clydesdales tours and live animal shows will be available for the 2015 season. Camel rides and parakeet feedings are also back by popular demand. As always, guests 21 and older can enjoy complimentary samples of Anheuser-Busch beers.
Grant's Farm will continue to offer once-daily Estate Walk Tours, which showcase the grounds of the Busch family estate while guides share details on the rich history of the property. Tours are available at 9 a.m. Tuesday through Saturday and at 9:30 a.m. on Sunday for $25 per person.
Almost a century ago, August A. Busch Jr.'s father purchased a piece of Missouri farmland once plowed and planted by Ulysses S. Grant. Today, the country estate of the Busch family, Grant's Farm, is preserved as a home for more than 1,000 exotic animals from six continents.
The beautiful oasis can be seen courtesy of Anheuser-Busch, Inc. An open-air Eisenbahn coach winds past Mirror Lake to Grant's Cabin, which was completed in 1856 and remains today as the only structure hand built by an American president. The fence across from the cabin was constructed from 2,563 rifle barrels as a Civil War memorial.
The coach travels through The Deer Park where more than 100 species of bison, elk and antelope to zebras, llamas and ostriches can be seen. You can pet and hand feed some of the animals and attend the entertaining, educational shows featuring African elephants, parrot and birds of prey at the Tier Garten Amphitheater. Admission is free. Bring your camera and take home special memories.
Grant's Farm is home to a breeding operation for the world-famous Budweiser Clydesdales. The tour features an opportunity to see the Clydesdale sires, mares and foals up-close in their pastures next to the Clydesdale Stable.
Admission to Grant's Farm, tram rides and all shows is complimentary.
Parking fees:
* $12 per vehicle
* $30 for buses, RV's and oversized vehicles
* $40 for a season parking pass, available at the General's Store
* Stroller rental available
* Wheelchair accessible.
For information on brewery tours, see Anheuser-Busch.
The Saint Louis Front Page is owned and maintained by the Moore Design Group for the sole purpose of disseminating news and information about the Metropolitan Saint Louis area. Text or graphics may not be copied, rewritten or distributed in any manner whatsoever without written permission. For more information, contact email@example.com All rights reserved world wide © 1996 - 2015 Moore Design Group.
Vacation Savings at
Drury Hotels
Call 1-800-325-0481
Fine Art Prints of St. Louis Attractions


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You can now invite anyone to meet at your favorite Starbucks.
Limited-time feature
53 year old man | Clarksville, TN, USA | Seeking women 40-55 within 50 miles
Courtesy is as much a mark of a gentleman as courage". Theodore Roosevelt
Thank you for reading my profile, 3 points I would like to make before wasting your time: 1. I would like to meet an equal In the sense that having independents is not an issue but to be embraced and supported. However If your all your free time revolves around the local bar please move on. 2. I would like to meet someone who's not dependent on any chemicals or alcohol to have a good time or "deal with life". I enjoy cocktails and wine like the next guy however the key word would be enjoy in that statement. 3. Live, Love, laugh and family should be the soul of one's life. Commitment to excellence in your career and health, playing hard and passion should be the foundation. If your good with all this with no drama; Let's rock! I have been told am an attractive Italian that's a great cook also. Any man that does not treat women with respect, genuine kindness and caring is absolutely is making a mistake. True Love can be given without being shared, but a shared Love still allows for your special loved one to grow. To me, there is nothing more special than a shared Love. Having a sense of humor and being able to enjoy a day playing flag football in the mud and dress for an elegant evening out on the town a few hours later for cocktails and dinner. Also enjoy as much a night of movies, wine, a good quilt and smiles : ). Who am I? I am a professional, home owner financially independent, Athletic, enjoys working out and health conscious. secure, career man, intelligent, goal minded, assure of himself and stable. Am looking for a friend / soul mate in hope to be my best friend in life. I enjoy places where there's entertainment of all kinds, laughs, friends, and no stress, Live music particularly jazz, playing softball, fine dining, wine tasting, movies, playing pool, ect, ect. Hope to find someone to share moments together and just be an oasis in each other's lives in this crazy world.
Aerobics, Baseball, Cycling, Walking / Hiking, Weights / Machines
I exercise 3-4 times per week.
I like Cats, Dogs
None right now...
Middle of the Road
That's the adventure…..Were we are is the place to be. Concert, plays, dinner theater, titans games or whatever.
Live music particularly jazz, playing softball, fine dining, wine tasting, movies, playing pool, life
Any type of competitive sports, entertainment I can share with my lady friend or with friends from racket ball, bowling to poker. I love to talk SMACK and enjoy. I love to travel, like the great tan in my profile photo, my girls were so jealous.
Replay by Ken Grimwood
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|5'5" (165cms)
|5'0" (152cms) to 8'5" (256cms)
|Body type:
|Athletic and toned
|Slender, About average
|No preference
|Salt and pepper
|No preference
|Social Drinker
|Social Drinker
|Executive / Management
|No preference
|I'll tell you later
|No preference
|Never Married, Widow / Widower, Currently Separated, Divorced
|Have kids:
|Yes, and they live away from home (More than 3)
|No preference
|Wants kids:
|No, but it's OK if my partner has kids
|No preference
|White / Caucasian
|No preference
|Christian / Catholic
|No preference
|Some college
|No preference


text:
Computing Facilities
448 and 475 Birge Hall
Open 24 hours (key access)
The Physics/Astronomy Network of Intel Computers (PANIC) cluster is a general purpose cluster of Linux computers for use by Physics faculty and unaffiliated graduate students. It consists of two computer labs with 9 PCs and two laser printers. In addition to the standard Linux applications, several third-party applications like Mathematica and R are installed, and users are provided with a small amount of network disk space for data storage. Individual office workstations can also be hooked up to the PANIC facility by request. Visit the PANIC website for more detailed information about this computer cluster and the instructions on how to obtain a PANIC account.
PANIC Support
PANIC is supported by the L&S Computer Resources Unix team.
448 and 475 Birge Hall
Open 24 hours (key access)
If you have a problem or request, please send email to: email@example.com
Dan Ackerman is the primary manager of the PANIC cluster:
154 New LeConte
(510) 219-7684
Public Printers
Learn more about the Physics Departments' public printers.
Most rooms in Birge and LeConte halls have 100Mbps ethernet connections. To find out your IP address or to request a new connection for your lab or office see Anthony Vitan, 151 LeConte Hall, 642-2242.
Wireless Access
Learn more about our wireless access.


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Is there an inside fuse box?
Ok well sorry for the noob question, i bought my ram last weekend and today finally brought it to the garage to do a few things. One thing i noticed, is there a fuse box inside? I went to hardwire my Valentine one, and couldnt find it...haha.
Im guessing the one under the hood is the only one?
Which leads me to my next question. Do they make extending wires for something that needs to be plugged into the fuze box.
For example:
I need to plug in that fuze for the Valentine 1 but deff wont reach.
This is how it looked in the last car:
Any idea's?
The answer to your first question, is "no". There isn't a fuse box inside the truck.
The answer to your second question is that you can extend the wire as long as you need to. You just need some wire, a pair of wire cutters, and a means of reconnecting the two ends. The best way to connect 'em is by soldering, but I tend to just use butt connectors, as they're a lot easier to use, and do a good job.
good luck
Thanks, was hoping there was one inside..lol
Hey, I'm probably too late with this information, but I noticed you said you were hardwiring your valentine one. I read about this company while I was searching the boards one day - mounts the detector to your rear view mirror, and then you can draw power from behind the mirror tapping into the connector back there. Pretty slick setup -
I might be wrong but isn't there a fuse box on the side of the dash? You have to have the driver door open to access it? My memory sucks though....maybe I'm thinking of my s10....
The 2nd generation Rams have a fuse box on the left side of the dash. This was discontinued in 2002. :(
Well, you have me on that one. I've never owned an S10 so can't say a single intelligent thing about one. :dork:
It is too bad that there are no convenient power connections in the interior of the trucks other then the cig lighter and/or power ports. Not everyone is OK with wires hanging all over the place. Funny thing; other then pure battery power, that are no convenient connections under the hood either. LOL
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text:
Nick Farley Photographer
Natalie221Model Female Brisbane, Queensland, Australia My MM URL: http://www.modelmayhem.com/naataaliiee
Mayhem # 2508488
About MeI've just recently had my first photo shoot, which was better than I'd expected. Before that I was a hand model for an international Nail Technician competition. Over the past few months i've lost 30kg, hoping to try modelling. Alot of people say I have a unique look, and encouraged me to try my hand at becoming a model. It has always been a dream career and lifestyle of mine, but now I'd like to make it happen.
By joining this site, I hope to eventually be noticed by someone who may offer me an opportunity. If you'd like to contact me, please send me a private message and I'll get back to you!
Nick Farley Photographer
ausfx Photographer
GAS Imaging Photographer
Jake Darwin Model
J Marini Photographer
Arachnie Model
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text:
Thanks to the support of active members in Germany, after the first Germany Roadshow in 2009, the OW2 Management Office is organizing for the second time in 2011 OW2 meetups in Germany. The objective of these short events is to meet potential partners and members and develop awareness about OW2. We are very grateful to the Fraunhofer Fokus Institute for hosting the workshop and for their support in the overall organization.
Fraunhofer Forum (directions)
Anna Louisa Karsch Straße 2
10178 Berlin
TENTATIVE AGENDA: (this agenda will be updated progressively)
10:00: Introduction and eGovernment@Fraunhofer FOKUS – Mr. Gerd Schürmann
10:15: OW2 overview and open source trends analysis - Cedric Thomas, OW2 CEO
10:45: Governmental institutions and Open Source - Speaker TBC
11:15: SMBs & national and international corporations - Speaker TBC: Microsoft Interop group
11:45: QualiPSo CC @ Fraunhofer FOKUS – Mr. Yuri Glickman
12:00: Lunch break
13:00: OW2 Technical activities overview - Alexandre Lefebvre, OW2 CTO & France Telecom R&D
13:20: Chameleon project: one year into OW2 (Clément Escoffier, Alice Murphy, Akquinet, A.G.)
13:40: JOnAS project (Jacques Cayuela, Bull)
14:00: Spago projects (Stefano Scamuzzo, Engineering)
14:20: Coffee break
14:30: SMBs national and international corporations 1
14:45: SMBs national and international corporations 2
15:00: Panel Discussion: Open Source Interoperability eGovernment
Useful Links
Community Events


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RIP Sherman Helmsley. First Ballot Inductee into the Black Walk Hall of Fame.
Jul 24, 2012 @ 5:00pm
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Sorry ladies, he's now Kim Jung Un-Available.
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Hearty Beef Hodgepodge
8 Jul 2009
I have learnt this Hearty Beef Hodgepodge from one of my friend. This Hearty Beef Hodgepodge delight should be tried by everyone and I am sure you will always crave for more!!
|Ground beef
|1 1⁄2 Pound
|Chopped onion
|1⁄4 Cup (4 tbs)
|1 Clove (5 gm), minced
|3 Cup (48 tbs)
|Condensed minestrone soup
|3 Can (30 oz)
|Tomato soup
|1 Can (10 oz)
|Canned pork and beans
|28 Ounce (1 Can)
|Chopped celery
|1 1⁄2 Cup (24 tbs)
|Worcestershire sauce
|1 Tablespoon
|1 Teaspoon
|Taco mix
|1 Tablespoon (1 Envelope)
Brown the ground beef with the onion and garlic in a skillet, stirring until crumbly; drain.
Combine the ground beef mixture, water, soups, pork and beans, celery, worcestershire sauce, oregano and taco mix in a large kettle; bring to a boil.
Reduce heat and simmer for 30 minutes.
Recipe Summary
Course:Side Dish
Method:Stir Fried
Rate It
0 Comments
Hearty Beef Hodgepodge Recipe


text: In Pine Valley, Adam finds out that Kendall was driving the car that ran Greenlee off of the road, Krystal is torn about telling David the truth and Kendall is prepared to say goodbye to Bianca! Across the way in Port Charles, though Nik opens up to Rebecca, she's eyeing Lucky, Anthony approaches Claudia, while Sonny and Jason are on the hunt for revenge! In Llanview, David and the Buchanans get a DNA shocker, Schuyler reaches out to Cole and Starr... » Read More


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Two More Images of CM7 on the G2X
Pictured to the left are pics of CM7 running on the G2X, courtesy of Chris Soyars. Again, this is still very much a work in progress, but we figured you would want to see some more good news if you are the owner of a G2X.
About author
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U.S. military policies today marginalize women and entrench the masculinization of political life at home and abroad.
Issues / War & Peace
Indonesias military buildup and East Timor-style militia activities threaten to destabilize Papua and the region.
Massive injections of U.S. and Soviet arms have kept the war raging between northern and southern Sudan for nearly a half-century.
Since Indonesias invasion of East Timor in 1975, the U.S. has supplied the Indonesian army with more than $1 billion in arms.
In the U.S. the attractions of missile defense endure, fueled most recently by the apparent Gulf War successes of the Patriot missiles and by perceived threats of long-range missile launches by so-called rogue states.
Drug crop eradication has produced little effect on the price or availability of cocaine in the United States.
As leaders of 34 Western Hemisphere countries gather in Quebec City, Canada in April 2001, President George W. Bush hopes that the third Summit of the Americas will mark a step toward fulfilling his fathers dream of creating a free trade area stretching from Alaska to Tierra del Fuego.
Given the atmosphere of suspicion and distrust that so often characterizes U.S.-China relations, it is vitally important that Chinese foreign policy and military capabilities be calmly and carefully assessed.
The Bush administration's defense review affords an opportunity to overhaul the nation's military strategy, forces, and equipment plans in light of the challenges and opportunities of the new century.
Violence and warfare in Colombia are often blamed on the drug trade, but their roots run much deeper and go back well over five decades.


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Simply confirm your registered email address below and click "Reset Password." We will immediately email you a link back to the site where you can enter a new password for this account.
The second winner of the Doritos Crash the Superbowl fan made commercial contest.
WRAT 95.9 FM The Rat Rocks!Your feedback is important! Click here to send us your comments or suggestions.


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A great choice for your 11 o'clock break.
Give your lunch time a kick with this spicy wrap
Delightful lunch time option made with Dubliner Cheese.
You're taste buds won't be able to resist the delicious smell while this panini toasts!
A classic pasta dish.
A perfect flavour combination!
Kids will love these fun sandwiches in their lunchboxes


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Otolaryngologist (ear, nose, throat)
48 years of experience
Accepting new patients
48 years of experience
Accepting new patients
Logan - Ogontz - Fern Rock
5501 Old York Rd
Philadelphia, PA 19141 Phone number & directions
5501 Old York Rd
Philadelphia, PA 19141 Phone number & directions
License & Identifications
A medical license is required for a doctor to practice medicine in a particular state. Requirements vary by state but most require, at a minimum, post-graduate training in the doctor's specific specialty. An NPI number is a national identifier unique to an individual health care provider and is managed by the Center for Medicare & Medicaid Services (CMS).
Learn more about Dr. Wayne Weddington:
View the full profile to get information on:
- Qualifications
- What's important to you
View full profile
Patient Reviews
Patient comments (0)
The overall average patient rating of Dr. Wayne Weddington is Excellent.
Specialties & Research
Dr. Wayne Weddington, MD is board certified in Otolaryngology.
- He has special expertise in 5 areas:
- Allergy
- Heart Failure
- Laser Surgery
- Otology
- Sleep Disorders
Education & Awards
Dr. Weddington graduated from Howard University , Washington.
Dr. Weddington has 7 hospital affiliations:
- Albert Einstein Medical Center
- Temple East, Inc.
Home > Find an Otolaryngologist > PA > Philadelphia Otolaryngologists > Dr. Wayne Weddington, MD
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- Additional Information
- Dr. Wayne P Weddington MD is a male Otolaryngologist, has 48 years of experience and practices in Otolaryngology.
- Where does Dr. Wayne P. Weddington practice?
- Dr. Wayne Weddington has 1 office location.
Dr. Wayne Weddington practices Otolaryngology near Philadelphia, PA.
See Philadelphia Otolaryngologists.


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We love traveling, but we don't love feeling cramped and uncomfortable. Try these yoga poses and stretches to counteract the negative effects of traveling.
Yoga can help us fight infections by boosting our immune system, reducing stress and strengthening our body's functions and systems.
Take your yoga practice to a whole new level. Check out one of the latest trends in the fitness world: TRX yoga. Here are three poses to help you get started.
Just because you finished your run doesn't mean you're done for the day. Try these yoga poses tailored to the needs of runners.
Belly bloat is a common part of life, even if you keep a healthy lifestyle. Flatten your stomach with these moves to help things pass through your body easily, like a cleanse.
Read More Yoga Articles
So you say you're not flexible? Join the club. These 12 poses are perfect to help you gain flexibility from head to toe. Give them a try.
More Yoga Tips, Training & Advice
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Page 1
Something from (almost) nothing:
the impact of multiple displacement
amplification on microbial ecology
Erik K Binga1, Roger S Lasken2and Josh D Neufeld1
1Department of Biology, University of Waterloo, Waterloo, Ontario, Canada and2J. Craig Venter Institute,
La Jolla, CA, USA
Microbial ecology is a field that applies molecular techniques to analyze genes and communities
associated with a plethora of unique environments on this planet. In the past, low biomass and the
predominance of a few abundant community members have impeded the application of techniques
such as PCR, microarray analysis and metagenomics to complex microbial populations. In the
absence of suitable cultivation methods, it was not possible to obtain DNA samples from individual
microorganisms. Recently, a method called multiple displacement amplification (MDA) has been
used to circumvent these limitations by amplifying DNA from microbial communities in low-biomass
environments, individual cells from uncultivated microbial species and active organisms obtained
through stable isotope probing incubations. This review describes the development and
applications of MDA, discusses its strengths and limitations and highlights the impact of MDA on
the field of microbial ecology. Whole genome amplification via MDA has increased access to the
genomic DNA of uncultivated microorganisms and low-biomass environments and represents a
'power tool' in the molecular toolbox of microbial ecologists.
The ISME Journal (2008) 2, 233–241; doi:10.1038/ismej.2008.10; published online 7 February 2008
Subject Category: microbial population and community ecology
Keywords: metagenomics; microbial ecology; multiple displacement amplification; phi29 DNA
polymerase; whole genome amplification; single-cell microbiology
Environmental microbial diversity is poorly under-
stood and the majority of microbes are inaccessible
by laboratory cultivation. As a result, microbial
ecology has sought to develop molecular methods to
characterize whole communities. The extraction of
community nucleic acids is a typical initial step,
and although cell biomass is often sufficiently high
to enable analysis by PCR, gene hybridization or
metagenomics, this is not always the case. Because
microbial ecologists study challenging environ-
ments, such as insect guts (Broderick et al., 2004),
ice cores (Christner et al., 2001), permafrost (Steven
et al., 2006), deep subsurface sediments (Teske,
2005) and air (Brodie et al., 2007), high-sensitivity
PCR protocols have been required for the analysis of
single genes. A whole genome amplification (WGA)
step provides access to community DNA from these
low-biomass environments. In addition, the vast
number of uncultivated organisms associated with
the 'rare biosphere' (Sogin et al., 2006) contain
enzyme-encoding genes that hold great promise for
medicine, biotechnology and industry. Fortunately
for microbial ecologists in the early 21st century,
small yields of DNA from low-biomass communities
or individual uncultivated cells are readily retrieved
by the advent of WGAvia the multiple displacement
amplification (MDA) reaction. Here we highlight the
history of MDA and discuss its mechanism, cap-
abilities and limitations. The applications of MDA
in microbial ecology are reviewed for accessing low-
abundance DNA from cells or environmental sam-
ples. Finally, we discuss the future of MDA and its
potential integration into additional facets of micro-
bial ecology research.
A brief history of MDA
Initial WGA reactions utilized PCR-based techni-
ques such as degenerate oligonucleotide primed
PCR (Telenius et al., 1992) and primer extension
PCR (Zhang et al., 1992). However, these were
limited by nonspecific artifacts of amplification
(Cheung and Nelson, 1996), strong bias (Paunio
et al., 1996) and short amplification products
(Telenius et al., 1992; Zhang et al., 1992; Paunio
Published online 7 February 2008
Correspondence: JD Neufeld, Department of Biology, University
of Waterloo, 200 University Avenue West, Waterloo, Ontario,
Canada N2L 3G1.
E-mail: firstname.lastname@example.com
The ISME Journal (2008) 2, 233–241
& 2008 International Society for Microbial Ecology All rights reserved 1751-7362/08 $30.00
Page 2
et al., 1996). MDA was the first WGA method based
on an isothermal reaction (Dean et al., 2001).
Double-stranded DNA template is initially dena-
tured before incubating at 301C for 2–16h, depend-
ing on the amount of starting template and the
commercial MDA kit being used (Table 1). MDA
utilizes target DNA template, buffer, dNTPs, random
polymerase, which is commonly derived from the
Bacillus subtilis bacteriophage phi29. The 30phos-
phorothioate modifications are required for amplifi-
cation, as they render the hexamers resistant to the
30–50exonuclease proofreading activity of the poly-
merase (Dean et al., 2001). The phi29 DNA poly-
merase extends the random primers and its strong
'strand displacement activity' allows it to displace
existing primer-originated extensions downstream
(Figure 1). Continued priming and strand displace-
ment generates a branched structure and gives
exponential DNA amplification. The reaction yields
double-stranded linear DNA, single-stranded forms
and some remaining branched intermediate struc-
tures (Figure 1). Complete denaturation of MDA
products and subsequent resolution on denaturing
alkaline agarose gels revealed a 12kb average
length of the resulting single-stranded amplified
DNA (Dean et al., 2002). The reaction is terminated
by heatingto 651C
the polymerase, and yields up to 40mg of DNA per
50-ml reaction.
The phi29 DNA polymerase was first isolated by
Blanco and Salas (1984) from Escherichia coli cells
expressing the P2 gene, which encodes the sole
phi29-associated DNA polymerase. It has an extre-
hexamers andDNA
for 10min,denaturing
mely high processivity, adding an average of 70000
nucleotides each time it binds the primer template
(Blanco et al., 1989). Use of single-stranded M13
DNA as template gave a rolling-circle mode of
replication, in which the polymerase repeatedly
copied around the circular template via its strand
displacement activity, yielding a product of con-
catenated M13 repeats. The DNA polymerase's
associated 30–50exonuclease proofreading activity
results in a low intrinsic error rate of 10?6–10?7
(Watabe et al., 1984; Blanco and Salas, 1985) and the
accumulation of mutations in MDA products at a
Table 1 Several commercially available kits for multiple displacement amplification of DNA
ProviderKit Intended template for amplificationIncubation
Volume (ml)Time (h)Yield (mg)
GE Healthcare
(Baie d'Urfe ´,
Quebec, Canada)
GE Healthcare
TempliPhi 100/500Small circular DNA (for example,
TempliPhi Large
TempliPhi HT
Large circular DNA (for example,
BACs, cosmids and fosmids)
Small circular DNA (for example,
Circular templates containing
'difficult' sequence (for example,
repeats, inverted sequences and
Linear DNA
Linear DNA (high yield)
Linear and circular DNA
GE Healthcare20183.5–4
GE HealthcareTempliPhi
Sequence Resolver
10 181
GE Healthcare
GE Healthcare
Ontario, Canada)
GenomiPhi V2
GenomiPhi HY
REPLI-g Mini
REPLI-g Ultrafast
REPLI-g Midi
Linear and circular DNA201–1.57–10
Wisconsin, USA)
Linear and circular DNA
Customizable amplification
of linear DNA
Abbreviation: ND: not defined in the manufacturer's protocol.
Figure 1
structures formed during multiple displacement amplification
(MDA). The arrowheads represent the location of the phi29
polymerase synthesizing DNA in a 50to 30direction.
Sequential diagrammatic representation of branched
Multiple displacement amplification and microbial ecology
EK Binga et al
The ISME Journal
Page 3
rate of only 10?5–10?6(Esteban et al., 1993; Nelson
et al., 2002), nearly 1000-fold less than for PCR
using the Taq DNA polymerase (Dunning et al.,
1988; Saiki et al., 1988).
The first use of MDA for amplifying whole
genomes targeted human DNA (Dean et al., 2002)
and as few as 90 copies of the genome (0.3ng DNA)
yielded more than 30mg of product. For eight loci
assessed, amplification bias (over- and underrepre-
sentation of sequences) was several orders of
magnitude less than for the PCR-based WGA (Dean
et al., 2002). Since then, MDA has been applied to
many more eukaryotic studies, including the geno-
mic sequencing and genotyping of humans and
primates (Lovmar et al., 2003; Barker et al., 2004;
Paez et al., 2004; Dickson et al., 2005; Jiang et al.,
2005; Lu et al., 2005; Ro ¨nn et al., 2006), insects
(Gorrochotegui-Escalante and Black, 2003), fungi
(Foster and Monahan, 2005; Gadkar and Rillig, 2005;
Ferna ´ndez-Ortun ˜o et al., 2007) and additional
applications in forensics and medicine (Hosono
et al., 2003; Lasken and Egholm, 2003; Barber and
Foran, 2006; Spits et al., 2006; Ballantyne et al.,
Although MDA has enabled genomic sequencing
from low concentrations of template nucleic acid,
remaining limitations include (a) nonspecific am-
plification derived from primer dimer formation or
contaminating DNA template, (b) formation of
chimeric DNA rearrangements and (c) representa-
tion bias. Exciting new research indicates the
possibility of improvements to the specificity,
fidelity and sensitivity of MDA. For example,
single-strand binding protein and spermidine were
reported to improve DNA yield up to 66%, with
reduced representational bias and increased sensi-
tivity for targeted templates (Wu et al., 2006).
Similar effects were reported with the mutant
single-strand binding protein protein of Thermus
thermophilus (Inoue et al., 2006). One report
suggested that background synthesis from primer–
primer interactions may also be reduced or elimi-
nated by the modification of hexamer primers with
the incorporation of one or two 50-terminal universal
bases (Lage et al., 2003). Recent work demonstrated
that carefully avoiding trace levels of DNA contam-
ination resulted in only 0.36–1.0% of sequences
being nonspecific (Marcy et al., 2007a).
The concentration and copy number of DNA
template affect the MDA reaction. Hutchison et al.
(2005) discovered that a smaller MDA reaction
volume (3ml) greatly reduced the amount of non-
specific DNA generated with 50 molecules of single-
stranded circular M13 template compared to higher
reaction volumes (15 or 30ml). Others have shown
that the total number of DNA templates affects
representational bias (Lasken et al., 2005), with a
single copy of template giving the highest bias
(Raghunathan et al., 2004). Reducing bacterial
DNA template from 10ng (4106genome copies) by
10- and 100–1000-fold reduced representational bias
3- and 6-fold, respectively (Wu et al., 2006). Other
studies also showed increased bias with decreased
template copy number (Detter et al., 2002; Bergen
et al., 2005; Holbrook et al., 2005; Kalyuzhnaya
et al., 2006; Neufeld et al., 2008).
Chimera formation and amplification bias are
concerns for genomic sequencing of DNA generated
from MDA. Zhang et al. (2006) discovered that
cloned MDA products included up to 50% chimeric
inserts (3kb average length). They postulated (in-
correctly) that chimeras were generated by the
cloning step, with E. coli rearranging the cloned
and branched DNA. Remarkably, chimeras were
reduced by 80% (to a final rate of 6% of inserts
screened) through three enzymatic treatments of (a)
phi29 DNA polymerase and dNTPs in the absence of
primers to 'debranch' the DNA, (b) S1 nuclease to
digest remaining single-stranded DNA present in the
mixture and (c) flow shearing and size selection on
agarose gels followed by nick translation with E. coli
DNA polymerase I.
The predominant mechanism for chimera forma-
tion has now been elucidated and occurs during the
MDA reaction rather than in the subsequent cloning
step (Lasken and Stockwell, 2007). Pyrosequencing
(454 Life Sciences method; a cloning-independent
approach) of DNA amplified from single E. coli cells
revealed a chimera frequency (one chimeric junction
per 22kb) similar to that previously detected (Zhang
et al., 2006) by cloning-dependent Sanger sequen-
cing. The principal mechanism of chimera forma-
tion was through initial extension of random
primers followed by displacement of these 30ends
and re-priming on a second template, often where
there was a short region of complimentarity of
2–21bp. The second templates were usually nearby
50ends such that 85% of chimeras consisted of two
segments joined in inverted orientation (Lasken and
Stockwell, 2007). It was suggested that after ampli-
fication, digestion with S1 nuclease (Zhang et al.,
2006) may be effective by digesting single-stranded
regions postulated to occur as MDA reaction inter-
mediates in the pathway leading to chimeras
(Lasken and Stockwell, 2007).
Use of MDA in microbial ecology
Single-cell amplification
Microbial ecology is in the early stages of establish-
ing links between uncultivated microorganisms in
the environment and their trophic roles within
communities (Neufeld et al., 2007). Progress has
been made in the ability to analyze genomes,
without the prerequisite of cultivation, by using
MDA from individual cells. The first demonstration
of genomic sequencing from single cells used E. coli
isolated by fluorescence-activated cell sorting, fol-
lowed by MDA (Raghunathan et al., 2004). Ampli-
fication of B5fg of template generated B24mg of
product, more than a billion-fold amplification. No
Multiple displacement amplification and microbial ecology
EK Binga et al
The ISME Journal
Page 4
miscalls were found in a 662-bp 16S rRNA gene
sequence amplified from MDA DNA, demonstrating
that cultivation was no longer a prerequisite for
genomic analysis. Owing
synthesis, only 30% of the amplified DNA was
actually E. coli genomic sequence. Recent work has
achieved X99% specific amplification from a single
cell (Marcy et al., 2007a), possibly due to more
stringent protocols for avoiding trace DNA contami-
nants. MDA was first used to sequence from an
uncultured cell isolated directly from the environ-
ment using fluorescence in situ hybridization (FISH)
of 16S rRNA to identify and capture soil microbes by
micromanipulation (Lasken et al., 2005). Microma-
nipulation is relatively time consuming but is a
powerful research tool allowing observation of cell
morphology, repeated washing in rinse buffers to
remove contaminating free DNA and visual docu-
mentation that a single cell was captured and
delivered to the MDA reaction (Ishøy et al., 2006;
Kvist et al., 2007). As discussed below, the isolation
of single or a few cells for use in MDA reactions has
also been carried out by fluorescence-activated cell
sorting (Raghunathan et al., 2004; Podar et al., 2007;
Stepanauskas and Sieracki, 2007), microfluidics
(Marcy et al., 2007a,b) or micromanipulation (Kvist
et al., 2007). Cell sorting by FISH labelling of 16S
rRNA followed by automated cell lysis and MDA
using liquid-handling stations is expected to pro-
vide unprecedented high throughput for the analy-
sis of uncultivated cells.
Single-cell analysis is complementary to metage-
nomics (Lasken, 2007), which uses bulk environ-
mental DNA samples to rapidly obtain genomic
sequence but can be limited by the difficulty of
assembling individual genomes from multiple or-
ganisms (Tringe et al., 2005; Rusch et al., 2007).
Even partial genomic drafts from single cells
provide genetically linked sequences useful for
guiding the assembly of metagenomic sequences
into discrete genomes (Lasken, 2007). In a proof of
principle for community analysis, cells from a
defined mixture of three methylotrophic bacteria
were separated by flow sorting (Kalyuzhnaya et al.,
2006) using 16S rRNA FISH probes that distin-
guished between these three methylotrophs. When
101, 102and 103cells were collected in each tube
and subjected to MDA, 20%, 50% and 95% of
resulting DNA sequences correctly associated with
the FISH-targeted organism, and none were from the
'contaminating' organisms; the remaining sequences
corresponded to nonspecific DNA synthesis gener-
ated as an artifact of MDA. All of the tested clones
represented the desired microbe when 104and 105
cells were used as template in MDA.
For environmental samples, the use of FISH
probes for 16S rRNA in fluorescence-activated cell
sorting prior to MDA was demonstrated by Podar
et al. (2007) targeting the uncultivated phylum TM7,
which existed at low relative abundance (p2%) in a
California soil sample. Amplification of DNA from
to nonspecific DNA
five sorted cells resulted in detection of the TM7 16S
rRNA gene, but also labelled sequences from
contaminating Pseudomonas spp DNA. Nonethe-
less, they collected 420000 sequences that as-
sembled into 132 contigs, encoding 670 predicted
TM7 proteins. The TM7 representatives were found
to share similarity to Chloroflexi and possess genes
associated with multidrug resistance, starvation
resistance and DNArestriction
Further, TM7 cells possibly employ a twitching
motility and may be involved in biofilm formation,
both suggestions supported by the retrieval of genes
encoding a Type IV pilus. Organisms from the TM7
phylum also occur at a low relative abundance
(p2%) on human subgingival tooth surfaces. Single
TM7 cells were examined in dental plaque commu-
nities with a FISH-targeted, custom-designed micro-
fluidic system coupled with two rounds of MDA
(Marcy et al., 2007b). Of the 2.86Mb sequence
data obtained from cloned DNA, 1474 genes were
identified across 288 scaffolds with X3 genes.
Several genes were associated with Type IV pilus
sequences, similar to those found in soil TM7 cells
(Podar et al., 2007). Together, these studies offered a
previously unavailable glimpse into the potential
metabolism and community function of this wide-
spread and uncultivated bacterial division.
In another study (Kvist et al., 2007), micromani-
pulation was used to retrieve the uncultivated clade
C1b crenarchaeotes from agricultural soil in Den-
mark. These may be major contributors to ammonia
oxidation in world soils (Leininger et al., 2006), yet
they have only been detected in agricultural soil
clone libraries (Ochsenreiter et al., 2003), a fosmid
metagenomic library from a soil sample (Quaiser
et al., 2002) and in root-extract enrichment cultures
(Simon et al., 2005). Cells were disrupted from the
soil matrix, isolated with a micromanipulation
method for individual FISH probe-positive cells
(Ishøy et al., 2006) and used for MDA and DNA
sequencing (Kvist et al., 2007). Clone libraries
confirmed that DNA from C1b phylotypes was
amplified for two of the eight collected cells.
Sequencing 97 plasmid clones yielded 130kb and
more than 60 open reading frames, 20 of which were
unambiguously associated with the collected cre-
narchaeote. The remaining clones were of unknown
origin, but were quite possibly associated with the
archaeal cells collected in this study. Notably, one of
the open reading frames closely matched a nitrous
oxide reductase from an uncultivated soil bacter-
ium, indicating that some soil crenarchaeotes may
be capable of dissimilatory nitrous oxide reduction.
As an alternative to sorting cells with fluores-
cence, cells or extracted genomic DNA from parti-
cular organisms may be amenable to alternative
purification methods prior to MDA. Mavingui et al.
(2005) purified cells and DNA of an obligate
intracellular pathogen (Wolbachia pipientis) from a
large amount of host material, using differential
centrifugation and pulsed-field gel electrophoresis.
Multiple displacement amplification and microbial ecology
EK Binga et al
The ISME Journal
Page 5
The B50ng of retrieved Wolbachia DNA was
amplified via MDA. Multiple PCR primer sets
confirmed that the MDA products preserved the
gene arrangements found in the original template. In
another study, individual filaments, each containing
several hundred cells of the large sulfur bacteria
Beggiatoa, were physically removed from contam-
inating sediment cells and subjected to MDA
(Mu?mann et al., 2007). Sequences representing an
estimated 70% of the genome were then obtained
with 454 pyrosequencing. The subsequent bioinfor-
matic analysis confirmed the chemolithoautotrophic
metabolism proposed for Beggiatoa in 1888, and
suggested that other electron donors and acceptors
may be coupled in this fascinating sulfur-oxidizing
Methods for isolating cells and characterizing
their genomes are expected to become increasingly
sophisticated and high throughput (Lasken, 2007;
Marcy et al., 2007a), especially methods that target
organisms associated with the 'rare biosphere'
(Sogin et al., 2006) and techniques specifically
linked to the incorporation of labelled substrates
(Huang et al., 2007). As a result, the use of MDA to
obtain genomic DNA from single cells is anticipated
to play a central and increasing role in accessing
genetic and ecological information associated with
uncultivated microorganisms.
Whole community genome amplification
Microbial ecologists have used DNA-based meth-
odologies to study the diversity and composition of
microbial communities in biomass-rich environ-
ments for the past two decades. However, the study
of low-biomass environments with PCR, DNA
hybridization (for example, microarrays and dot-
blot hybridization) or metagenomics has been
limited by contaminating humic acids (for PCR)
and low yields of extracted nucleic acid for
community analyses (for PCR, hybridization and
metagenomics). Metagenomics provides a broad
view of a community's genetic composition, but
requires microgram quantities of DNA to construct
libraries for screening or sequencing. While ex-
tracted DNA may not be limiting from surface soils
or sediments, it may be limiting from environments
of low bacterial abundance. Whole community
genome amplification using MDA has recently
overcome these limitations, enabling the molecular
assessment of microbial communities that would
not have been accessible otherwise.
One difficulty for low-biomass environments is
that associated humic acids and exopolysaccharides
may inhibit PCR-based analysis. To alleviate this,
Gonzalez et al. (2005) used MDA as a pre-PCR
enrichment step on samples from a cave, meadow
soil and a wastewater treatment system. MDA from a
defined mixture of pure-culture DNA accurately
reflected the unamplified mixture based on denatur-
ing gradient gel electrophoresis fingerprints. An
analysis of Wolbachia DNA from infected mites
(Jeyaprakash and Hoy, 2004) and a study of
Mycobacterium leprae from human skin samples
(Groathouse et al., 2006) have also demonstrated
MDA as a powerful preamplification step when PCR
is hindered by low template concentration or
coextracted cellular PCR inhibitors.
In an example of a community analysis using
MDA, microorganisms associated with a lace coral
(Pocillopora damicornis) were studied (Yokouchi
et al., 2006). Limited sample sizes and the difficulty
of extracting quantifiable DNA from coral made this
an ideal community to access through MDA. First,
MDA representational bias was assessed using
cyanobacterium Synechocystis sp PCC6803. Ten
single-copy genes assayed by quantitative PCR (after
MDA) were all amplified between 104and 105times
by MDA, a substantial improvement over degenerate
oligonucleotide primed PCR and primer extension
PCR, which had amplification bias over a range of
3–6 orders of magnitude for tested sequences (Dean
et al., 2002). To analyze the coral microbial commu-
nity, 0.4ng of extracted DNAwas subjected to MDA,
yielding 24mg of X10-kb product. Approximately
70% of sequences were in common between PCR-
generated 16S rRNA gene libraries derived from
MDA and control (unamplified) DNA, indicating
that MDA maintains high representation of commu-
nity composition. The study demonstrated that
Proteobacteria (Alphaproteobacteria in particular)
were dominant phylotypes in this coral DNA
Subsurface and contaminated soil microbial po-
pulations are examples of communities in which
metagenomic library construction would be diffi-
cult. Abulencia et al. (2006) estimated that an
untenable 11–88kg of contaminated subsurface soil
(at 104cellsg–1) would be necessary to extract
microgram amounts of DNA for shotgun library
construction, with 20-fold more required for BAC
libraries. To overcome this, MDA was successfully
applied to heavily contaminated subsurface-soil
samples from Oak Ridge, Tennessee, to generate
sufficient template for 16S rRNA gene PCR and
construction of libraries. Prior to MDA, only one of
three samples gave useful libraries. Diversity esti-
mates also indicated that the libraries from ampli-
fied DNA extracts possessed a higher level of
diversity. Small-insert metagenomic libraries gener-
ated from three MDA reactions shared a similar
cluster of orthologous group profiles, indicating
conservation of sequences by MDA. However,
MDA from several microbes, experimentally mixed
in known ratios, indicated that some bias may occur
in their representation (Abulencia et al., 2006). In
another subsurface study, Edwards et al. (2006)
demonstrated the first use of 454 pyrosequencing to
characterize metagenomic DNA, employing MDA to
obtain sufficient template using microbial assem-
blages from the Soudan Iron Mine in Minnesota,
USA. Pyrosequencing of MDA-amplified microbial
Multiple displacement amplification and microbial ecology
EK Binga et al
The ISME Journal


text:
This Week's Front Page
Amanda Bachelder teaches crafting classes at Camas Antiques using Annie Sloan Chalk Paint. Some of the items she has painted include her boots and lunch box, in addition to furniture. She also has a space at the store which features antiques, collectibles, romantic home decor and vintage inspired handmade goods.
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More BlackBerry Z10 images leak
It's the BlackBerry Z10 in its full glory as more and more images keep surfacing on the Internet. Feast your eyes, it's only 32 days away. I can't wait to get my hands on it and if this happens to be the final product I'm sold and I love it.


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Asparagus and Bok-Choy Frittata
Asian sesame oil and ginger add an aromatic twist to this simple, yet sophisticated egg dish.
Level: Moderate
Serves: 4
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- 2 tbsp. Cooking oil
- 3 scallions including green tops
- 1 tsp. grated fresh ginger
- 1 clove garlic
- 1 small head bok choy (about 3/4 pound)
- 0.75 lb. asparagus
- 0.75 tsp. salt
- 9 eggs
- 0.25 tsp. fresh-ground black pepper
- 1 tsp. Asian sesame oil
- Heat the oven to 325°F. In a medium cast-iron or ovenproof nonstick frying pan, heat the cooking oil over moderate heat. Add the scallions, ginger and garlic and cook, stirring, until fragrant, about 30 seconds. Add the bok choy and cook, stirring, until the leaves wilt, about 2 minutes. Add the asparagus and 1/2 teaspoon of the salt and continue to cook, stirring occasionally, until the vegetables are almost tender, about 3 minutes more.
- Evenly distribute the vegetables in the pan and then add the eggs, pepper and the remaining 1/4 teaspoon of salt. Cook the frittata, without stirring, until the edges start to set, about 2 minutes. Put the frittata in the oven and bake until firm, about 25 minutes. Drizzle the sesame oil over the top.
- Notes: If you've been avoiding eggs for fear of their high cholesterol content, there's good news: The latest scientific research shows overwhelming evidence that saturated fat, not dietary cholesterol, is what can affect blood cholesterol. Eggs are nutrient dense. In fact, they're the highest-quality source of protein available (after mother's milk) and they're even low in calories (seventy-five per large egg). So for almost everyone, eating an egg or two every day is perfectly fine.
- Wine Recommendation: South Africa is one of the so-called New World wine countries, along with Australia, New Zealand, and the Americas, but its wines best reflect the balance of the European tradition. A South African chardonnay will make a marvelous partner for this Italian-inspired dish.
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The UGG Classic Cardy is a heathered merino wool boot made to look like your favorite sweater. Three wooden buttons etched with the signature UGG logo allow this boot to be one of UGG's most versatile styles. Slouch the boot down, cuff it all the way over or wear it all the way up. All boots in the UGG Classic Collection feature a soft foam insole covered with genuine sheepskin and have a molded EVA light and flexible outsole designed for amazing comfort with every step.
Ugg Classic Cardy Women's Boot
Features include:
This style tends to run a size large. We recommend ordering a size smaller from your typical shoe size. If you are between sizes, we recommend ordering a half size down.
I wear various Ugg Styles, but if you want to dress them up and show the detail of the whole boot, this one is EXCELLENT with either tights or leggings, and an above the knee skirt.
I'm only 5'4, so the tall styles tend to swallow up too much of my leg and make me look like a stump. ...but then the short styles are too casual no matter what you wear with them...
So, this is by far the best looking of the UGG styles if you yourself are not on the tall side, but want to wear an Ugg that is!
I love these cardys as much as i like my classics. I also like how you can wear them three different ways, that is what attracted me to them in the first place!
Super cute boot that is very comfortable- I usually wear a size larger but they recomended ordering a size smaller and they are a perfect fit.I love the different options for wearing this boot- I am over 6 feet tall and yet they come up high enough if I want to wear them up and I like them folded over as well as slouched down.
These boots are the best! I love the legwarmer look of the sweater knit. Perfect for a cold day to keep those toes warm! :-)
My only recommendation is to get your true size. I normally wear an 8.5, but because of the recommedation on the product page to get a size smaller, I ordered an 8. It was too little, and I had to exchange for a 9.
I wear these at work and at play. They are so comfortable and stylish. I've had many compliments. I love that the Grey is not too dark and not too light. This is my 2nd pair of Uggs. I love these more than the original. Try a pair. You won't be sorry you did.
This is a great Ugg boot. Very stylish and can be worn so many different ways! The grey is so beautiful and goes with everything. The only downfall is that sometimes its hard to keep the boot up on your calf. It tends to like to slouch down a lot even when the top button is fastened. Otherwise a great comfortbale buy from a quality brand.
I wear them at home as well as outside. Very warm; perfect for NYC's cold days.
I love these shoes because they go with pretty much anything. And I love how you can wear them different ways to go with the look youre going for.
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David Anderson, Lonsdale, Minn. letter: Legal, illegal immigration are not the same
Published 08/26/2013, Grand Forks Herald
Shelton Gunaratne wants Herald readers to believe that illegal and legal immigration are the same and serve the national interest ("Immigration serves U.S. economy and culture," column, Page A4, Aug. 20). He could not be more wrong.Word count: 388
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I think that this is long overdue, but........
since disney owns the muppets, isnt it about time that the muppet characters start making their appearances in the parks??
I think that this is long overdue, but........
since disney owns the muppets, isnt it about time that the muppet characters start making their appearances in the parks??
A Muppet parade would be cool as well as an update to the Muppet attraction.
Sig by Poisonedapples!
I agree.
I've seen Kermit in Disney's MGM(or is it Pixar yet?) Studios in WDW. For autographs he had a stamp to put in the kids' books. They could do that at DCA.
A decent attraction is needed. The 3D film is iight the first few times but I rather see a Roger Rabbit type "Muppet Mayhem" kind of attraction. I wouldn't mind if they knocked down Muppet 3D in place of an attraction.
Yeah, the kids would like to meet the frog.
A stamp for an autograph would be fine too.
I agree and definitely would love to see more Muppets in DCA. I'd love to see a parade, characters and/or an updated attraction.
IMO, The Muppets and Star Wars both need expanded presences within Disney parks.
I've seen Miss Piggy walking the park. She's kinda' creepy. Like she's 6 feet tall and huge!
How come it is always me who ends up on the 'other' side of things?
**checks page 313 of 'Dianetics', muttering, "What's wrong with me?"**
Here it goes: I was never in love with non-Disney franchises having prominent representation in the Parks. I certainly realize that the Muppets have been 'acquired' by Disney. These characters however are famous as non-Disney entities.
More reliance on non-Disney entertainment seems a poor precedent to set. The 'Star Wars' franchise is still a Lucas property, more the reason to limit its influence in the Parks. Would it be cool to have Kermit and Vader running around the Park? Maybe, but ultimately I believe it is a dangerous foundation to build upon thematically.
Last edited by fo'c's'le swab; 08-14-2007 at 08:41 AM.
i've also seen miss piggy walk around, but i think this would be a great idea. i'd actually like to see more characters in general at dca.
Going for Halloween 2011
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I don't mind em in DCA.... however, If I see em in DL I'm gonna take Kermit into NOS and Have me some frog legs.
All your dreams can come true if you have the courage to pursue them. - Walt Disney
I kinda' agree with you Fo. I think first Disney needs to really put out better quality movies, in order for those characters to be represented in the part. The last few years have really lacked and they've relied on Pixar too much. Because their movies have been lacking, character recognition isn't high. I don't know how Disney can fix the problem, except write better scripts, don't rely so much on computer animation, and write characters that resonate with kids.
That being said: I am a huge Muppets fan. I love the Fraggles. Big Bird was my first crush. Oh! And I am a 2nd generation Muppets fan (my dad looks like Honey Dew, but loves Beaker.) so if I could see the Muppets (not the 6ft tall Piggy, but a normal sized Gonzo... like a real puppet sized one), I'd be so starstruck. The Muppets 3D lacks though.