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Heath Ledger's Daughter Matilda Will Inherit Her Father's 'Dark Knight' Oscar
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2016 MTV Movie Awards - Hottest Girls on The Carpet
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Henry Cavill's Got Some Stiff Competition! The Rock Shows Off His Superhero Package At The MTV Movie Awards
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The stunning and hilarious star won the Comedic Genius award at the MTV movie awards and looked great in a black jumpsuit.Genevieve Van Voorhis
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Pregnant, cant sleep well, body temperature is also high. Will it effect the baby ?
Hi, my wife is pregnant for 8 months. She told me she can t sleep well cos she has to get up 3 or 4 times during night sleeping. Actually, She normally is fine but these few days she s not well. Last night her temp is higher or she feels to get a fever . Hmmm, should we go to see the doctor?........whether does it affect the baby?....... how to handle with the fever?......... Thank you so much in advance for your kind advise and recommendation :)
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Sat, 16 Feb 2013
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Dr. Aarti Abraham
Mar 2013
Thank you for your query.
After the 8th month of
, due to various reasons, sleep it disturbed at night.
It could be frequent urge to urinate, and slight postural discomfort and breathlessness due to the growing tummy (
Ask her to sleep in propped up position or left lateral, in a rocking chair if it comfortable, adn to make up her sleep during the daytime.
Measure the temperature, and if it is higher than 100.4 degrees, please consult your doctor.
Take care.
Answered: Thu, 21 Feb 2013
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Thank you for your query.
After the 8th month of pregnancy, due to various reasons, sleep it disturbed at night.
It could be frequent urge to urinate, and slight postural discomfort and breathlessness due to the growing tummy ( uterus ).
Ask her to sleep in propped up position or left lateral, in a rocking chair if it comfortable, adn to make up her sleep during the daytime.
Measure the temperature, and if it is higher than 100.4 degrees, please consult your doctor.
Take care.
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subset: commoncrawl


text:
Hacked By: selectLOL
Hack Information:
Keyhacks: Press 1 Toggle health - 2 Medipacks
Game Information
Parody of classical fantasy role-playing games with three playable characters and amusing battle system.
Cheats: Hack Menu: 12 Gun Buttons - Points +500 - All Guns 50 XP - H...
Hacked By: DPETTY
Cheats: Hackbar: Press [1] Fairy - [2] Key - [3] Bomb - [4] Fire mod...
Hacked By: selectLOL
Cheats: Hackbar: Press [1] Next Level - [2] Previous Level
Hacked By: leverage419
Cheats: Hackbar: Press [Z] Cash - [X] Golden Spheres - [C] Win Level
Hacked By: leverage419
totally not a major reference to the colour of magic and/or the light fantastic
rock paper scissors shoot *Shoots enemy*
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the elves are link
Plus the battle system is lame, it is just rock, paper, scissors.
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Registered Users: 477038
Hacks Played Today: 115396
Total Hacks Played: 813381992
There are 1117 users online
26 Members, 1091 Guests


text:
An asinine adventure through the world of high fantasy.4.42 / 5.00 1,432 Views
Nothing To Fear
Super villainy at its worst ...4.17 / 5.00 2,987 Views
The Ascent of Alfred
Make Alfred Great Again4.11 / 5.00 6,905 Views
loving it. :D hope u make stickman can't fight 6 :D
already a work in progress
haha its so funny !!
thank you
different is with 2 F !!!
alfalfa is too! :D
wht the fck was that??!!
it was the greatest stick figure animation of 2012
It's not bad. It's pretty well drawn, funny, and with memes! The bad thing is, the sound kinda sucks and you misspelled some stuff.
yeah. i need to learn to get better quality audio out of these movies. and i know my spelling is terible (< on purpose) but i'll proof-read everything for the next one ;)


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ABC Responds to Nicollette Sheridan's Lawsuit
ABC has spoken about the lawsuit filed by Nicollette Sheridan against the network and "Desperate Housewives" creator Marc Cherry. In its statement to ET, the Alphabet said they have not seen the actual complaint but have recalled similar claims made by the actress last year.
Sheridan said in the lawsuit that Cherry was "behaving in an extremely abusive and aggressive manner" while on the set of the series. She pointed out one incident in 2008 where Cherry pulled her aside and struck her in the head as well as the face after she questioned something about the script. When Sheridan decided to report it to ABC execs, they did nothing significant to address the situation and Cherry later wrote off her character.
In its responding statement ABC said, "While we have yet to see the actual complaint, we investigated similar claims made by Ms. Sheridan last year and found them to be without merit."
Nicollette Sheridan who is seeking $20 million in compensation, is suing for wrongful termination, and assault and battery, among others. Marc Cherry still remains mum about the case.
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- For accurate prices please enter the date of your trip!
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Our best offer
196.82 € / person / night, without provision
Description of the accommodation:
Rooms, services When taking a trip to Newport Beach, booking a room at The Fairmont Newport Beach may be a great decision. The hotel is offering 444 tastefully furnished rooms to those seeking accommodation in Newport Beach. In case you'd like to have a meal or a drink on the spot, you will be provided freshly prepared dishes in the restaurant every day. For guests arriving by car parking possibility is available nearby. Relaxing wellness services will be available to you at the hotel. You may also rent a car to discover all must-sees of the area. The fans of actively spent leisure time may take the nearby opportunities to rent a bike. You can also try the local gym and its provided services. Airport transfer can be requested upon preliminary notice. The bar has excellent atmosphere for a relaxed evening talk or a personal meeting. In the safe your valuables may be the safely secured. You can browse the Internet via wireless connection (WiFi) available at the hotel. The exact list of services available at The Fairmont Newport Beach (including those for an extra fee) and the exact list of equipment in a room can be found on the information sheet below.
Payment For the reservation you may need to provide advance payment by credit card or credit card guarantee, depending on the exact date, room and price.
Arrival, opening hours The earliest check-in time is 16:00, the latest check-out time is 12:00. The personnel can speak English.
The Fairmont Newport Beach foglalás, szoba árak Foglalás, szoba árak
- For accurate prices please enter the date of your trip!
Accommodation services
Parking (32 €/night)
WiFi in shared spaces (10 €/night)
Relaxation: Fitness, Bike rental at the accommodation
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Default services: Pet friendly (50 €/night), Bar, Elevator, Safe deposit box
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News & Politics
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We tease out the complex history behind those baffling events in the news.
October 26th, 2014
Episode 38 of 130 episodes
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I've got my website secured via passwords setup thru NT User Manager.<BR>After the user logs in, I'd like to capture the usename and store it to a cookie to track the user thruout the site. How can I do this?
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1st – Frankie Wainman Junior
2nd – Rob Speak
3rd – Lee Fairhurst
1st – Mick Sworder
2nd – Paul Harrison
3rd – Mark Gilbank
The World Championship is run over a series of qualifying rounds (see the fixture list). The top 48 qualifiers are split into two semi-final rounds. The semi-final grid is lined up different to a normal BriSCA F1 race, with the highest points scorers at the front. The top 10 finishers from each semi-final go through to the World Final. Again drivers are seeded on the grid with the highest qualifiers on the front row.
The winner of the World Final gets the honour of wearing a Gold Roof on their car for the following 12 months.
Visit the World Championship Hall of Fame for a list of past winners.
1st – Frankie Wainman Junior
2nd – Rob Speak
3rd – Lee Fairhurst
1st – Mick Sworder
2nd – Paul Harrison
3rd – Mark Gilbank
Here is the low down on the overseas qualifiers for the 2016 World Final.
Before next weekend's World Championship at Coventry, Scott Reeves takes a look at the drivers to watch out for.
Nigel Green took the top spot in the World Championship Semi-Final, while Chris Cowley dominated the meeting final.
Rob Speak claimed the first spot on the front row for the World Final in September, while Bobby Griffin took his second Sheffield meeting final of the season.
It's nearly Semi Final time! Here's who to look out for at Sheffield and Buxton.
The BSCDA has announced the official World Semi-final grids for the 2016 BriSCA F1 World Championship
Before next weekend's World Championship, Scott Reeves takes a look at the drivers to look out for.
Rob Speak dominated the World Championship Semi-Final, but it was Michael Steward who won the meeting final.
Davidson dominated the heats, but it was Nigel Green who took the meeting final at Shootout Round #5.
Rob Speak made it three finals in a row
Rob Speak took his second final win in a row in Shootout Round 3 at King's Lynn.
Photo gallery from Shootout Round #2 at Skegness
Our next unsung hero is EdCreations' Eddie Cooper
Mick Sworder took the first final win of the 2016 Shootout.
The 2016 World Championship race
Michael Scriven won the World Masters at Northampton


text:
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BMW Concept X5 eDrive to Make North American Debut at NYIAS
Related: 2014 BMW X5 Trims and Specs
Sometimes, two things just go together perfectly: Chocolate and peanut butter, sriracha and everything, Kim and Yeezy. BMW has gathered up two of its technologies—eDrive and xDrive—and brought them together under one sexy roof. That roof, belonging to the BMW Concept X5 eDrive, will make sweet harmony for the first time this week at the New York International Auto Show 2014.
The BMW Concept X5 eDrive and its slightly unwieldy name look to wow the assembled masses in New York with performance that is anything but cumbersome. The concept utilizes a drive system with a 245-horsepower four-cylinder engine mated to a 95-horsepower electric motor powered by a lithium-ion battery, generating the power needed to get the most out of BMW's xDrive four-wheel drive system and the efficiency of intelligent hybrid technology that allows travel of up to 19 miles on a single charge without using a single drop of fuel.
While the BMW Concept X5 eDrive is making its North American debut, it is not an entirely new model: the X5 eDrive originally debuted at the 2013 Frankfurt Motor Show, and it appears that very little has changed in the time since apart from some minute aesthetic details. What may have potentially changed in the time since its first appearance is BMW's stance on bringing a production version to the masses. Is it possible that the Spartanburg, South Carolina BMW plant, which was recently the recipient of a $1 billion investment, could house production for a newer and more efficient addition to the X5 lineup? Perhaps we'll find out more in the days to come.
Related: The 2014 BMW X5: Success Breeds Success
For more information on the BMW Concept X5 eDrive, check out BMW's press release. For the latest news out of New York, stop back by The News Wheel this week!
Brought to you by BMW of Santa Maria also serving San Luis Obispo, Santa Barbara, and Lompoc, California.


text:
Princess Party Cookbook
Over 100 Delicious Recipes and Fun Ideas
by Annabel Karmel
- #5437 in Nonfiction, Food & Drink
- #1857 in Nonfiction, Family & Relationships, Parenting
- #1018 in Nonfiction, Health & Well Being, Health, Healthy Living
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Every little girl dreams of being a princess . . .
And there's nothing a princess likes more than throwing a party! This enchanting new book allows you to indulge the princess in your little girl with recipes and ideas designed to feed her imagination and encourage her participation in the kitchen.
From Fairy Finger Sandwiches and Rice Krispies Hearts to Emerald Pizza, Ruby-Glazed Chicken, and Gelatin Gems, these are magical, delicious dishes fit for a royal banquet. You'll be able to celebrate every occasion in style, with Snowflake Cupcakes for the holidays, Pretzel Broomsticks for Halloween, and Tropical Smoothies for the beach.
Filled with ideas for creating themed invitations and table settings, plus tips for making jewelry, gifts, and decorations, this is truly the indispensable guide to the perfect princess party!
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by on October 22, 2016
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Laser and LED Specialist Omicron presents a new Generation of LED modules
The innovative team of Omicron has developed new high-performance LED modules. The updated product in comparison to the previous version is characterized by greater functionality and flexibility for applications in industry and research.
With more than 40 different wavelengths from deep UV to the near infrared and optical output power of several hundred milliwatts, the so-called "LEDMOD" series can be used in many applications such as microscopy, chemical analysis, spectroscopy, forensics and other areas.
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The new "LEDMOD high power" modules by Omicron were presented in February 2015 at the trade fair "Photonics West" in San Francisco (United States).
Further information on Omicron laser products can be found at www.omicron-laser.de
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Since 1989, Omicron has been developing, building and producing innovative laser systems. With a highly qualified team Omicron specialized in customized solutions for applications in the fields of medicine, research, biotechnology, such as microscopy and flow cytometry, digital imaging and optical data storage as well as quality assurance and measurement engineering. Product development and production comply with European and US guidelines. A broad band of laser sources in the range of UV VIS/IR is available to satisfy individual customer requirements. Omicron offers single light sources as well as complete system solutions. Omicron pursues the objective of being an industry leader in product development and has not only set trends in laser technology but also has drawn worldwide attention with its developments.
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Further reports about: > Flexibility > LED > LED modules > Omicron > USB > data storage > different wavelengths > laser technology > near infrared > optical data > optical data storage > product development
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In a paper in Scientific Reports, a research team at Worcester Polytechnic Institute describes a novel light-activated phenomenon that could become the basis for applications as diverse as microscopic robotic grippers and more efficient solar cells.
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By forcefully embedding two silicon atoms in a diamond matrix, Sandia researchers have demonstrated for the first time on a single chip all the components needed to create a quantum bridge to link quantum computers together.
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'Ferroelectric' materials can switch between different states of electrical polarization in response to an external electric field. This flexibility means they show promise for many applications, for example in electronic devices and computer memory. Current ferroelectric materials are highly valued for their thermal and chemical stability and rapid electro-mechanical responses, but creating a material that is scalable down to the tiny sizes needed for technologies like silicon-based semiconductors (Si-based CMOS) has proven challenging.
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text:
Low back pain is a very common problem that is increasingly being treated surgically. This book aims to evaluate carefully the possible surgical approaches to low back pain, with detailed appraisal of the factors leading to their success or failure. It begins by explaining the scientific basis for surgery and considering the different diagnostic techniques that may be employed, thereby elucidating the surgical rationale, indications, and contraindications. The value of conservative options is also assessed to help the reader weigh the need for surgery. The various surgical modalities, including the most recent, are then fully described and evaluated with the aid of numerous illustrations. The book concludes with a chapter devoted to evidence-based analysis of the outcome of surgery in patients with low back pain. This book will be invaluable to orthopaedic and neurosurgeons, rheumatologists, neurologists, and all who are concerned with the effective treatment of this often debilitating condition.... 65 3 Biomechanics 65 3 Gynecology/obstetrics 55 2 Psychology 55 2 Research methods 50 2 Clinical pediatrics and geriatrics 50 2 First aid and ... The contrast between the two programs is dramatic in the area of clinical clerkships, which averaged 3, 467h in medicine vs. ... sciences, which consists of extensive laboratory and hands-on training in manual procedures and has no equivalent in medicine.
|Surgery for Low Back Pain
|Marek Szpalski, Robert Gunzburg, Björn L. Rydevik, Jean-Charles Le Huec, Michael Mayer
|Springer Science & Business Media - 2010-03-26


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If you log in, you can comment on buildings, submit new photos or update photos that you've already submitted.
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Description: 11 and 13, the Square
Grade: II
Date Listed: 16 May 1988
English Heritage Building ID: 164724
OS Grid Reference: TA0334026377
OS Grid Coordinates: 503340, 426377
Latitude/Longitude: 53.7236, -0.4353
Location: 4 Southgate, Hessle, East Riding of Yorkshire HU13 0AF
Locality: Hessle
County: East Riding of Yorkshire
Country: England
Postcode: HU13 0AF
Incorrect location/postcode? Submit a correction!
(north side)
9/79 11/13 The Square
House and shops. Mid C18 with later alterations. Brick, colour-washed,
pantiled roofs. L-shaped range of 2 storeys, 3 bays, with C20 2-storey
single-bay extension to right. 2 storeys, 6 bays to Southgate elevation.
Ground floor: C20 shop fronts to left with C20 corner porch with Ionic
columns. To right, a 16-pane sash with sill under cambered wedge lintel.
Elliptical arch with wrought-iron gates to C20 extension. First floor: four
16-pane sashes with sills under wedge lintels. Southgate elevation has C20
shop fronts to ground floor and 5 irregularly spaced 16-pane sashes with
sills under wedge lintels to first floor, similar 12-pane sash between left-
hand sashes. End and axial stacks, hip at junction of ranges. Includes
frontage to Southgate (Numbers 6, 8 and 10) (east side).
Listing NGR: TA0334226379
This text is a legacy record and has not been updated since the building was originally listed. Details of the building may have changed in the intervening time. You should not rely on this listing as an accurate description of the building.
Source: English Heritage
Listed building text is © Crown Copyright. Reproduced under licence.


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You are here: Home page Living & Sleeping Entertainment & relaxing baby walker Pinolino Lauflernwagen Auto Jonas
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Pinolino learning trolley car JonasPart of solid: Beech and MDF, brightly painted, L 54 cm, B 33 cm, 59 cm, Height of the handlebar 3-way adjustable from 59 to 64 cm, Rubber tyres, steel axles, Recommended for children from 1 to 6 years, Note: The doll is not More information > >
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Risk management solutions provider $AIZ continued to execute its multi-year transformation strategy in 1Q16. The sale of Employee Benefits to SunLife Financial closed on March 1. The wind-down of the health operations also remains on track. The membership continues to decline to only 26,000 covered lives as of March 31.
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Watching $TSLA Oct 28th $215 Calls for possible entry


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Home » Travel & Local » KaoshiungMetro
KaoshiungMetro - FREE
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Package name: com.gasolin.android.metro.kaoshiung , Last Update: 2012-11-30 02:11:31
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It's always happy hour when you consult your Spirit Guide. Your best friend for bar-hopping lets you in on the drink and food specials, tells you where the music is (and isn't) and helps you find the perfect spot. A great night out starts with Spirit Guide.
Chow's Asian Bistro
(505) 471-7120www.mychows.com
Mon-Sat: 11am-2:30pm, 5-9:30pm Sun: noon-2:30pm, 5-9pm (hours can change without notice)
Cuisine: Chinese
Chow's Asian BistroChow's is one of those rare Chinese food finds that doesn't make you feel too guilty about digging into a steaming pile of noodles. In fact, this place features extremely fresh and flavorful dishes that jump off the plate with color. Try the dragon sesame chicken and the volcano fish, not to mention their delicious herbal tea and gingered bok choy. Pick up some chopsticks and head to Chow's for lighter, yet zesty, fare. (Alibi Staff, August 11, 2010)
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Learn about the complexities of public representation and contestation among ideological factions in the Balkans. Runs through 10/31.
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Otto Fretter-Pico
From Wikipedia, the free encyclopedia
|The topic of this article may not meet Wikipedia's general notability guideline. (August 2016) (Learn how and when to remove this template message)
|Otto Fretter-Pico
|2 February 1893
Karlsruhe, Grand Duchy of Baden, German Empire
|30 July 1966
(aged 73)
Flims, Canton of Graubünden, Switzerland
| German Empire (to 1918)
Weimar Republic (to 1933)
Nazi Germany
|Years of service
|Major General (Generalleutnant)
|Commands held
|57th Infantry Division
148th Reserve Division
148th Infantry Division
World War II
|Maximilian Fretter-Pico (brother)
Otto Ludwig Julius Fritz Franz Balduin Fretter-Pico (2 February 1893 – 30 July 1966) was a highly decorated major-general (Generalleutnant) in the Wehrmacht during World War II who commanded several divisions. He was a recipient of the Knight's Cross of the Iron Cross, awarded by Nazi Germany to recognise extreme battlefield bravery or successful military leadership. Fretter-Pico surrendered on 28 April 1945 to Allied troops. After his release in July 1948, he settled in Switzerland. Fretter-Pico died at the age of 73 on July 30, 1966 in Flims, in the Canton of Graubünden, in Switzerland.
Awards and decorations[edit]
- Iron Cross (1914)
- Wound Badge (1914) in Black (fall 1918)[1]
- Knight's Cross 2nd Class of the Order of the Zähringer Lion with Swords (13 November 1916)[1]
- Honor Cross 3rd Class of the Royal House Order of Hohenzollern with Swords (April 1918)[1]
- Honour Cross of the World War 1914/1918 (21 December 1934)[1]
- Clasp to the Iron Cross (1939)
- General Assault Badge in Silver (30 September 1941)[1]
- German Cross in Gold on 11 December 1941 as Oberst in Artillerie-Regiment 297[2]
- Eastern Front Medal (11 August 1942)[1]
- Knight's Cross of the Iron Cross on 12 December 1944 as Generalleutnant and commander of 148th Infantry Division[3]
- Gassend, Jean-Loup (2014). Autopsy of a Battle, the Allied Liberation of the French Riviera, August September 1944. Atglen PA: Schiffer. ISBN 9780764345807.
- Patzwall, Klaus D.; Scherzer, Veit (2001). Das Deutsche Kreuz 1941 – 1945 Geschichte und Inhaber Band II [The German Cross 1941 – 1945 History and Recipients Volume 2] (in German). Norderstedt, Germany: Verlag Klaus D. Patzwall. ISBN 978-3-931533-45-8.
- Scherzer, Veit (2007). Die Ritterkreuzträger 1939–1945 Die Inhaber des Ritterkreuzes des Eisernen Kreuzes 1939 von Heer, Luftwaffe, Kriegsmarine, Waffen-SS, Volkssturm sowie mit Deutschland verbündeter Streitkräfte nach den Unterlagen des Bundesarchives [The Knight's Cross Bearers 1939–1945 The Holders of the Knight's Cross of the Iron Cross 1939 by Army, Air Force, Navy, Waffen-SS, Volkssturm and Allied Forces with Germany According to the Documents of the Federal Archives] (in German). Jena, Germany: Scherzers Miltaer-Verlag. ISBN 978-3-938845-17-2.
- Wegmann, Günter (2010). Die Ritterkreuzträger der Deutschen Wehrmacht 1939–1945 Teil III: Infanterie Band 7: Fl–Fu [The Knight's Cross Bearers of the German Wehrmacht 1939–1945 Part III: Infantry Volume 7: Fl–Fu] (in German). Osnabrück, Germany: Biblio-Verlag. ISBN 978-3-7648-2380-1.
|Military offices
|Preceded by
Generalleutnant Friedrich Siebert
|Commander of 57th Infantry Division
20 February 1943 – 1 September 1943
|Succeeded by
Generalleutnant Vincenz Müller
|Preceded by
Generalleutnant Friedrich-Wilhelm von Rothkirch und Panthen
|Commander of 148th Reserve Division
25 September 1943 – 20 March 1944
|Succeeded by
Generalleutnant Otto Schönherr
|Preceded by
Generalleutnant Otto Schönherr
|Commander of 148th Infantry Division
18 September 1944 – 28 April 1945
|Succeeded by
- 1893 births
- 1966 deaths
- People from Karlsruhe
- Lieutenant generals of the German Army (Wehrmacht)
- German military personnel of World War I
- Knights of the House Order of Hohenzollern
- Knights Second Class of the Order of the Zähringer Lion
- Recipients of the Gold German Cross
- Recipients of the Knight's Cross of the Iron Cross
- German prisoners of war
- People from the Grand Duchy of Baden
- Recipients of the clasp to the Iron Cross, 1st class
- German emigrants to Switzerland


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Oil and Acrylic on Canvas.
This artwork was made just after the november 15 Paris Attacks. When they catch him at last, the train explose on the city bridge. Feel like a Gotham air. Then, at this moment when too much deads appear.
Thanks for visiting my ArtWork. Why not seeing more?
All my artworks are delivered with a Certificate of Authenticity and Signed.
© 2016 Bast Etxeverria a.k.a. Etxebast. All rights reserved.
The images in context are not to scale and are for display purposes only. I have done my best to adjust the size of the painting as realistically as possible into the background picture.
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Lawmakers need to move forward in improving the Affordable Care Act, not backward in repealing it ...
White Acceptance Will Never Lead to Wealth in the Black Community
With jumi trillion in spending power, coupled with trillions in intellectual capital, Black people in this country can do anything we...
NAACP Portland Branch Meets May 28
The NAACP Portland branch invites the community to its monthly general membership meeting from noon to 2 p.m. May 28 at the American Red Cross Oregon Chapter, 3131 N. Vancouver Avenue. The meeting is open to all current and prospective members of the NAACP. President Jo Ann Hardesty and executive leadership will provide updates about current and future initiatives. This is an opportunity to connect with Portland NAACP's work in advancing racial justice in the community.
The mission of the National Association for the Advancement of Colored People is to ensure the political, educational, social, and economic equality of rights of all persons and to eliminate racial hatred and racial discrimination. The vision of the NAACP is to ensure a society in which all individuals have equal rights and there is no racial hatred or racial discrimination.
For more information about the NAACP Portland Branch, visit www.portlandnaacp1120.org, www.facebook.com/PDXNAACP or email email@example.com.
CAT to Present Renters Rights Workshops
The Community Alliance of Tenants will present a series of renters' rights workshops at Multnomah County Library locations in May.
The workshops will seek to educate tenants on their rights and responsibilities at all stages of the renting process: searching for a home, filling out applications, paying deposits and fees, getting repairs made, moving out and more. All workshops are first come, first served.
Media Artists Invited to Apply for RACC's 2016 Fellowship Award
The Regional Arts & Culture Council (RACC) is now accepting applications from media artists (including film, video and audio) for RACC's annual artist fellowship award. Applications are now available through the RACC GrantsOnline system at http://racc.culturegrants.org.
Since 1999, the RACC fellowship award has honored and supported uniquely talented local artists who contribute to the community in very meaningful ways. This year RACC plans to award two fellowships, with each artist receiving a cash award of $20,000. RACC rotates the recognition among four disciplines every year – visual arts, media arts, literature, and performing arts. Past Media Arts Fellows have included Jim Blashfield (2001), Chel White (2004), Joanna Priestley (2007) and Lawrence Johnson (2012).
Guidelines and applications are available in RACC's GrantsOnline system. To be considered, applicants must submit an Intent to Apply form no later than 5:00 pm, July 6, 2016.
A panel of community representatives with expertise in the media arts reflecting the disciplines of the applicants will select the two fellowship winners. An artist's involvement in the community will play a significant role in evaluating each application. In addition, applicants must meet several strict criteria in order to be eligible for these highly competitive awards:
Other RACC fellows are listed at racc.org/fellows, and they include: Mary Oslund, Obo Addy, Christine Bourdette, Terry Toedtemeier, Michele Glazer, Tomas Svoboda, Keith Scales, Judy Cooke, Michael Brophy, Craig Lesley, Thara Memory, Henk Pander, Kim Stafford, Robin Lane, Eric Stotik, Sallie Tisdale, Linda Austin, Anita Menon, David Eckard, and Ellen Lesperance.
For more information about the fellowship award and other RACC grants visit racc.org/grants.
Workforce Plan Open for 30-Day Public Comment
The Southwest Washington Workforce Development Council (SWWDC) is accepting public comment on its draft Strategic Workforce Plan for 2016-2020. Once finalized, the new plan will guide regional solutions for workforce development.
The plan is available for review at www.swwdc.org for 30 days beginning May 2.
Businesses, labor organizations, institutions of higher education, primary stakeholders and the general public are encouraged to provide comment on how SWWDC might better create the workforce solutions needed for our regional economy.
Comments should be made by emailing firstname.lastname@example.com by 5 p.m. Pacific Time on May 31.
For more Portland and Seattle events, see the Community Calendar.


text:
|Publication number
|Publication type
|Application number
|Publication date
|Jul 20, 1993
|Filing date
|May 4, 1992
|Priority date
|Dec 20, 1990
|Fee status
|Publication number
|Robert L. Brumley, Jr., Lloyd M. Smith
|Original Assignee
|Wisconsin Alumni Research Foundation
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (25), Non-Patent Citations (7), Referenced by (26), Classifications (6), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
This is a division of application Ser. No. 07/630,675, filed Dec. 20, 1990.
A. Field of the Invention
The present invention relates to the preparation and use of electrophoresis gels. The invention makes use of ultra-thin slab gels for the separation of macromolecules.
B. Description of the Art
Electrophoresis is the process of separating molecules on the basis of the molecule's migration in an electric field. In an electric field, a molecule will migrate towards the pole that carries a charge opposite to the charge carried by the molecule. The charge carried by a molecule often depends upon the pH of the medium in which the molecule is migrating. A common electrophoretic procedure is to set up solutions at different pH at each end of an electric field. At a certain pH, the isoelectric point of a molecule is obtained and the molecule carries no net charge. Therefore, as the molecule crosses the pH gradient, the molecule reaches an isoelectric point and is immobile in the electric field. This procedure separates molecules according to their different isoelectric points.
Electrophoresis in a polymeric gel, such as polyacrylamide or agarose, adds two advantages to an electrophoretic system. First, the polymeric gel stabilizes the electrophoretic system against convective disturbances. Second, the polymeric gel provides a porous passageway through which the molecules travel. Since larger molecules will travel more slowly through the passageways than smaller molecules, use of a polymeric gel permits the separation of molecules by either molecular size or isoelectric point.
Electrophoresis in a polymeric gel is also often used to separate molecules only by molecular size. Some groups of molecules, such as RNA and DNA molecules, all have the same electrophoretic mobility in free solution. These groups of molecules when moved through a polymeric gel by an electric field will segregate on the basis of molecular size. Thus nucleic acids and other groups of molecules with similar isoelectric points will migrate through the gel to be segregated solely on the basis of molecular size.
A polymeric gel electrophoresis system is typically set up in the following way: A gel-forming solution is allowed to polymerize between two glass plates that are held apart on two sides by spacers. These spacers determine the thickness of the gel. Sample wells are formed by inserting a comb-shaped mold into the liquid between the glass plates at one end and allowing the liquid to polymerize around the mold. The top and bottom of the polymerized gel are in electrical contact with two buffer reservoirs. Macromolecule samples are loaded into the sample wells. An electric field is set up across the gel, and the molecules begin to separate according to their size.
The size-sorted molecules can be visualized in several ways. After electrophoresis, the gels can be bathed in a DNA-specific or protein-specific stain which renders the groups of size-sorted molecules visible to the eye. For greater sensitivity, the molecules can be radioactively labelled and the gel exposed to X-ray film. The developed X-ray film will indicate the migration positions of the labelled molecules.
Both vertical and horizontal assemblies are routinely used in gel electrophoresis. The molecules can also be detected during electrophoresis, either by means of their intrinsic absorptive or fluorescent properties, or by labelling them with a detectable chromophore or fluorophore, or by other detection methods known in the art. In a vertical apparatus, the sample wells are formed in the same plane as the gel and are loaded vertically. The wells can be as deep and wide as needed, but the thickness of the well is limited by the thickness of the gel. If ultra-thin (<0.15 mm) gels are cast, loading the sample can be troublesome.
Ultra-thin electrophoretic gels are useful because they may be electrophoresed at a higher voltage. Therefore, the electrophoretic run is faster. Ultra-thin often gels yield higher resolution. Because of their thinness, the gels are fixed for autoradiography quickly and easily.
Sample wells in a horizontal apparatus are typically formed into the thickness of the gel and are loaded vertically. The wells may be of any desired thickness and, hence, are easier to load than the wells formed in a vertical apparatus. The depth of the wells is limited by the thickness of the horizontal gel.
The use of horizontal assemblies is known in the art. For example, Hurd, et al., U.S. Pat. Nos. 4,909,977 and 4,795,591, claim such a horizontal apparatus. The sample wells of the Hurd apparatus are formed by a comb-shaped mold at the extreme end of the slab gel. The comb is placed into a slot formed between the side of the bottom tray and the edge of the top tray.
Several problems are experienced by prior horizontal electrophoresis assemblies. The comb is held in place by the pressure of the top tray, but unless the comb is held very tightly, non-uniform sample wells with exogenous gel material will be formed. Additionally, the sample well geometry is such that the electric field "turns a corner" in the area of the sample well. This non-uniform electric field geometry will cause artifactual migration in this area of the gel and may cause the samples to electrophorese aberrantly. Finally, the apparatus is suitable to cast gels of 0.15-0.3 mm thick, so there is no teaching of the special problems of ultra-thin sample well formation.
The usefulness of electrophoresis depends on the sharp resolution of sample separation. This sharp resolution depends, in part, on the manner in which the macromolecular sample migrates from the sample well. Both the sample well and the electric field influence the migration of the macromolecules. Ideally, the sample wells would have a uniform, sharply defined size and have no extraneous pieces of polymerized gel that would interfere with sample migration. Uniform well size is necessary because the separation of molecules is often compared between samples that are electrophoresed side-by-side. Extraneous material in the sample well will cause impeded migration for part of the sample. Non-uniform sample migration greatly hinders high resolution molecular separation.
The placement of the electric field relative to the sample well is important because a non-uniform field can create artifactual results. Ideally, the electric field experienced by the loaded samples would be in a plane parallel to the gel, even when the sample is in the sample well. Then the molecules would experience a uniform electric field during their entire electrophoretic separation.
What is needed is an apparatus and method for casting ultra-thin gels having sample wells with a sharply defined, uniform geometry and having sample wells positioned so that the electric field passing through the sample well is parallel to the plane of the slab gel.
The present invention is an assembly for casting a slab gel and electrophoresing macromolecules in a slab gel. The assembly includes a horizontal base, a bottom plate bearing against the base, a top plate shorter than the bottom plate, with the top plate bearing on the bottom plate, an end plate bearing on the bottom plate at the loading end of the bottom plate, with the face of the end plate positioned relative to the face of the top plate so as to leave a gap that will admit a well-forming comb, a means for sealing the outer perimeter of the bottom plate and the top and end plates, a means for spacing the bottom plate from the top and end plates, a means for biasing the end plate against the top plate through a well-forming comb, and a means for passing electric current through a polymerized gel formed in the space created between the bottom plate and the top and end plates.
The present invention is also such an assembly in which the means for passing electric current through the polymerized gel formed in the space created between the bottom plate and the top and end plates includes an electrode with electrical contact to the polymerized gel through a gap through which an electric field originates. This assembly also includes a separate gap to permit sample loading through which an electric field does not originate.
The present invention is also a method for casting electrophoretic slab gels, using either assembly described above, including the steps of introducing sufficient gel-forming liquid between the top and end plates and the bottom plate to form a slab gel, inserting a well-forming comb into the gap created between the end plate and the top plate, biasing the end plate against the top plate through the comb so that the comb is held between the end plate and the top plate in a tight fit, permitting the gel-forming liquid to set, and removing the comb.
It is an object of the present invention to provide a method and apparatus that allow high resolution separation of macromolecules.
It is another object of the present invention to provide a method and apparatus capable of forming sharply defined, uniform samples wells in ultra-thin gels.
It is another object of the present invention to provide a method and apparatus capable of positioning sample wells so that the electric field passing through the sample well is parallel to the plane of the slab gel.
Other objects, advantages and features of the present invention will become apparent from the following specification when read in conjunction with the accompanying figures.
FIG. 1 is an exploded side view of a first preferred embodiment of the present invention.
FIG. 2 is a top view of the base and elements attached to the base of the first preferred embodiment of the present invention.
FIG. 3 is an exploded top view of the elements of the gel mold of the first embodiment of the present invention.
FIG. 4 is an exploded side view of the second preferred embodiment of the present invention.
FIG. 5 is a top view of the base and elements attached to the base of the second preferred embodiment of the present invention.
FIG. 6 is an exploded top view of the elements of the gel mold of the second preferred embodiment of the present invention.
FIG. 7 is a cross-sectional view of an assembled gel forming cavity of the embodiment of FIGS. 4 to 6.
FIGS. 1, 2 and 3 depict the structure of the first preferred embodiment of the present invention. The following description refers to these four figures.
The apparatus of FIG. 1 includes a large rectangular horizontal base 12 which has formed in its interior a large cavity or water jacket 14. The water jacket 14 is formed as a trough, open to the top, with its three lateral sides bounded by the base 12. Coolant may flow into and out of the water jacket 14 through inlet and outlet pipes 16 connected to the water jacket 14 through manifolds 18. The inlet and outlet pipes 16 are connected by appropriate tubing to a temperature controlled water reservoir, which is also equipped with a pump (not shown) which can circulate water through the water jacket 14. An elastomeric O-Ring 20 is fitted into a groove formed in the top of the base 10 extending around the outside of the trough of the water jacket 14. Four adjustable screwed legs 22 are threaded into bores at each corner of the base 12 to support the apparatus. An end alignment bar 24 is fixed in position attached to one end of the top of the base 12. Two guide blocks 26 are attached to the top of one side of the base 12.
FIGS. 1 and 3 illustrate the parts of the gel mold 28, which is formed of a bottom plate 30, a gel gasket 32, a top plate 34, two identical end plates 36 and four identical gap formers 38. FIG. 1 shows the pieces of the gel mold 28 in the order of their assembly. The gasket 32 is placed around the outer edge of the bottom plate 30. The thickness of the gasket 32 defines the thickness of the resulting gel. Gels which will usually be less than 150 μm, and may be as thin as 25 μm, may be formed by the present invention.
In assembling the parts of the gel mold, the top plate 34 is placed on top of the gasket 32 and the end plates 36 are also placed on the gasket 32 at each end of the top plate 34. In FIG. 3, 37 and 35 indicate the faces of the end plate 36 and the top plate 34, respectively which define the edges of the sample wells. In this embodiment, the end plate 36 is approximately one-sixth the length of the top plate 34, but the relative size relationship of these parts are not critical as long as the end plate is large enough to accommodate the agarose blocks. Gap formers 38 are inserted at each corner of the top plate 34 between the ends of the top plate 34 and the end plates 36. The assembled gel mold 28 forms a water tight reservoir for gel-forming liquid. A comb 46 of the same width as the gap formers 38 can then be fit into the gap formed between the edge of the top plate 34 and the edge 37 of the end plate 36.
The assembled gel mold 28 is positioned on the base 12 to abut the end alignment bar 24 and resting against the guide blocks 26. The gel mold 28 is secured tightly against the base 12 with a series of six clamps 40. The clamps 40 are placed around the sides of the base 12 and press onto the gel mold 28 by means of screws 42 threaded vertically through each clamp 40. Rotation of the screws 42 causes the top of the clamps to bear against the gel mold 28 to hold the mold 28 tightly in place against the base, thereby forming a fluid-tight seal between the lower surface of the bottom plate 30 and the O-ring 20.
At the sample-loading end of the mold 28 is an assembly for biasing the end plate 36 towards the top plate 34, against the gap formers 28 and the comb 46. In this embodiment, the assembly includes two pressure adjustment screws 48 threaded through the end alignment bar 24 which serve to apply horizontal pressure against the end plate 36.
Buffer chambers 50 are secured at either end of the mold 28 by four clamps 52. Screws 54 are threaded vertically through each clamp 52. Rotation of the screws 54 causes the top of the clamps 52 to bear against the top of the buffer chamber 50 to tightly hold the chamber against the top of the gel mold 28. An electrode holder 56 is attached to each buffer chamber 50. An adjustable screw 58 is threaded into the electrode holder 56 and secures and adjusts the carbon electrode 60. Not shown in the figures is the means for connecting a source of electric potential to the electrodes 60.
The end plates 36, top plate 34 and bottom plate 30 are preferably made of glass although other rigid transparent materials could be used such as quartz. Glass is preferred because it is electrically non-conductive and can be ground and polished to specific tolerances. If the gels are to be read optically, the glass should be one that has low fluorescence characteristics, such as fused silica glass. To ensure flatness of the glass components, optical quality glass, e.g. BK-7, Tempax, or soda-lime glass preferably polished to within 1 μm of flatness is used. The glass pieces are cut such that all sides are parallel. This parallelism is helpful for the proper alignment of the gel mold 28 on the base 12 once the components are assembled. The face 37 and 35 of the end plates 36 and the top plate 34 are polished flat to within about 5 μm. This uniform surface is helpful for the comb 46 to fit properly between the plate faces to form properly uniform wells.
The gap formers 38 and comb 46 are typically made of 0.030 inch sheet high density polyethylene. The base and attached elements may be formed of any suitable durable lightweight material, such as a dense plastic resin material.
The apparatus is used as follows. First all glass pieces are washed with a mild cleaner, rinsed with deionized water and thoroughly cleaned with three ethanol wipes. If the gels are to be fixed and dried on the bottom plate 30 after electrophoresis, the surface of the bottom glass is treated with gamma-methacryloxy-propyltrimethoxysilane which serves to bind the polyacrylamide to the glass surface. The top plate 34 may be siliconized to aid in the flow of polyacrylamide during gel pouring. Spacer surfaces may be coated with petroleum jelly to deter leakage. The gel mold 28 is assembled and positioned on the base 12 to abut the end alignment bar 24 and rest against the guide blocks 26. The screwed legs 22 positioned around the corners of the base are adjusted to provide a flat base 12. The gel mold 28 is secured onto the base with the six clamps 40. The buffer chambers 50 are clamped onto each end of the mold 28.
In this embodiment, the left-hand end of the mold 28 as viewed in FIG. 1 is the loading end. Gel-forming liquid, typically acrylamide, is introduced into the feed end buffer chamber 50 and allowed to flow into the empty space inside the gel mold 28. Enough liquid is introduced so that the liquid fills up the gel mold 28 and flows at each end into the gaps formed between the end plates 36 and the top plate 34. The gel-forming liquid is also introduced into the other chamber 50 when it has flowed across the cavity.
The well-forming comb 46 is inserted into the liquid-filled gap at the loading end. The bottom of the teeth of the comb 46 rest on the top of the bottom plate 30. The pressure adjustment screws 48 are twisted so that they bear against the end plate 36 and cause the comb 46 to be tightly squeezed between the end plate 36 and the top plate 34. Because of the tight fit of the comb 46, the leakage of gel-forming liquid around the comb 46 is strictly controlled. The wells thus formed are uniform, rectangular, sharply defined, and free of extraneous polymer that would impede electrophoresis or form irregularly shaped wells.
After the gel polymerizes, the comb 46 is carefully removed, leaving sample wells behind. The depth of these wells is determined by the length of the teeth of the comb 46. The sample wells may be formed up to the top of the top plate 34 and the end plate 36.
A small amount (approx. 1 ml.) of buffer is introduced into each buffer chamber 50. The water is also squirted with gentle pressure into the sample wells to remove residual gel-forming liquid and urea. The cooling water is begun circulating through the water jacket 14.
A 1 inch�1 inch�3 inch block of agarose, indicated at 62, (typically, 1.5% agarose in 1X TBE) is placed in each buffer chamber 50 with a small amount of buffer. The graphite rod electrodes 60 are positioned to be in contact with the top of the agarose blocks. Samples are loaded into the individual sample wells with a long needle syringe or pipette.
Voltage is applied across the electrophoretic slab gel, i.e. between the electrodes 60, in a short burst, typically 5000 volts for 6 seconds. During this short pulse, some of the sample moves into the slab gel. The voltage supply is turned off, and the sample wells are rinsed of excess sample. Electrophoretic resolution is improved because the migration of sample into the wells has been defined. After the wells are completely flushed, voltage is once again applied across the slab gel and the samples are electrophoresed for the desired period of time.
During electrophoresis, the gel temperature is continuously regulated by circulating coolant through the water jacket 14 under the gel mold 28. As shown in FIG. 1, coolant in the water jacket 14 will contact directly against the bottom plate 30. Coolant is dispersed across the width of the mold 28 by the manifolds 18. Coolant water is circulated by the pump associated with the external water reservoir (not shown). Because the gel is cooled while electrophoresis is taking place, up to 200 volts/centimeter of gel may be applied. Normally voltages of this intensity cannot be used in electrophoresis due to degradation of the polymeric gel from heat build-up in the gel. The cooling effect of the water jacket 14 prevents that here. A typical electrophoretic run for DNA sequencing procedures in a 75 μm 6% acrylamide gel is performed at 5000 volts for 20 minutes.
FIGS. 4, 5 and 6 depict the structure of the second preferred embodiment of the present invention. The following description refers to FIGS. 4, 5 and 6.
A horizontal base 112, a water jacket 114, a coolant inlet and outlet pipes 116, and inlet and outlet manifolds 118, height adjustment screw legs 120, mold clamps 122, mold clamp screws 124, buffer chamber clamps 126, buffer chamber clamp screws 128, and guide blocks 130 are identical structurally and functionally with those of the first embodiment and thus need not be described again in detail. An end alignment bar 132 is attached to the loading end of the base 112. This end alignment bar 132 has two pressure adjustment screws 134 threaded through it, as in the first embodiment. The end adjustment bar in this embodiment also holds the electrode assembly adjustment screws 136 which adjusts the position of the graphite electrode 138.
The gel mold 140 in this embodiment is defined by the surfaces of a bottom plate 142, and gasket 144, a top plate 146, a sample well and electrode assembly 148 and an electrode assembly chamber 150. FIGS. 4 and 6 depict these elements. The thickness of the gasket 144 defines the thickness of the gel. Gels as thin as 25 μm may be formed using this embodiment. The bottom plate 142 and the gasket 144 are identical structurally and functionally with those of the first embodiment. The top plate 146 has a lateral bar 152 attached to its top, at both its ends.
The sample well and electrode assembly 148, depicted in FIGS. 4 and 6, includes an assembly frame 153 which borders the assembly and divides the interior of the assembly into two open-topped chambers, a field transfer chamber 154 and a sample chamber 156. The field transfer chamber 154 has a plate 158 partially extending across its bottom. The plate 158 is positioned so that a gap at its front end permits electrical contact between material contained within the chamber 154 and the polymerized gel. The sample chamber 156, which is also open toward its front side, also has a plate 160 extending part of the way across its bottom. When the gel mold 140 is assembled, the lateral bar 152 forms the front side of the sample chamber. The plate 160 is positioned to leave a gap between the bottom of the sample chamber 156 and the edge top glass 146. This gap is the same width as the well-forming comb 162. The edge of the plate 160 and of the top glass 146 are polished to within 5 micrometers to provide accurate wells. A partition 164 separates the field transfer chamber 154 and the sample chamber 156. Note that the side edges of the sample well assembly frame 153 extend forwardly past the plate 160, or downward as illustrated in FIG. 5. The distance that the edges of the assembly frame 153 extends past the plate 160 is the width of the comb 162.
The electrode chamber 150 also has a plate 165 as its bottom. This plate 165 is positioned to leave a small gap for electrical contact and field transfer to the polymerized gel from the buffer chamber 150. An electrode assembly support 168 is attached to the base 112 at the non-loading end of the gel and holds the electrode adjustment screws 136 which support and position the electrode 138 at the non-loading end of the base 112.
Elements of the second embodiment are typically constructed of similar materials as in the first embodiment. The bottom plate 142, top plate 146 and the plates 158, 160 and 165 are preferably formed of glass. The frame of the sample well assembly 148 and buffer chamber 150 may be made of other materials, such as a durable plastic resin.
The second embodiment functions as follows. The top surface of the bottom plate 142 is treated for adhesion to the gel. The gel mold 140 is then assembled and positioned onto the base to fit against the guide blocks 130 and the end alignment bar 132, and is then secured onto the base 112 by the mold clamps 122 and the buffer chamber clamps 126. Height adjustment screw legs 120 are used to position the base 112 so that it is perfectly horizontal.
Gel-forming liquid is introduced into any of the gaps in the gel mold 140, such as the gap in front of the plate 158 in the field transfer chamber 154. Sufficient liquid is introduced to fill the mold 140 and the gaps. The comb 162 is inserted into the gap in the sample chamber 156 of the sample well assembly 148 and the pressure adjustment screws 134 are twisted so that the sample well assembly 148 is pressed against the top glass 146 with the comb 162 in place. The comb 162 is thus held tightly between the plate 160 that forms the bottom of the open-sided chamber and the top glass 146. This tight fit results in sharply defined, uniform wells.
The gel is allowed to polymerize, the pressure adjustment screws 134 are loosened, and the comb is carefully removed. The wells are flushed with distilled water. Excess water is wiped away from the plate 160 and the bottom of the sample chamber 156 with a tissue. A minute amount of water is left in each sample well.
In this embodiment, the electrodes are contacted directly to an extension of the gel itself. After the gel-forming material has polymerized in the gel mold, a second batch of gel-forming material is formulated. While a typical polyacrylamide gel is made by mixing 10 ml 6% polyacrylamide with 50 microliters of 10% APS and 5 microliters TEMED, the second batch of 10 ml of gel-forming solution material is made with three times the concentration of TEMED initiator. This second batch of gel-forming material is poured into the charge transfer chambers 154 and 150, and allowed to polymerize for approximately two hours. The result is a gel that extends continuously from one charge transfer chamber to the other through the gel mold 140. The electrodes impose the field on the gel by being placed in direct physical contact with the gels formed in the charge transfer chambers. Thus no buffer or other intermediary material is required to transfer the electric field into the gel.
In this embodiment, the electrical contact takes place through the field transfer chamber 154 and not through the sample wells. The electric field travels from the electrodes, through the gel in the charge transfer chambers, through the gaps formed adjacent plates 160 and 165 and then horizontally through the gel. Thus although the path of the electric field is bent in the chambers and in passing through the gaps, in the entire length of the gel in which the electrophoresis is performed, the field is linearly horizontal. Also, when the electric field passes through the sample wells, it is in a linear plane approximately parallel to the slab gel. Samples electrophoresed in this manner will not suffer from a period of artifactual migration that occurs when the electric field "turns a corner," as it does in the first embodiment and many prior art horizontal gel devices. This improved electric field geometry results in sharper, more uniform sample resolution and separation.
It has been discovered that the electrophoresis apparatus constructed in accordance with the present invention is capable of accurately, efficiently and quickly resolving DNA samples, particularly for DNA sequencing procedures. The provision for the base plate 142, which is continuously cooled on its underside, enables a thin polyacrylamide gel to be made and used without temperature build-up. Because higher voltages can be used, the separation of DNA strands can be accomplished more rapidly than previously possible, with no loss of accuracy or resolution.
These two embodiments do not represent the full scope of the invention. The invention may be employed in other embodiments, as well. Therefore, reference should be made to the claims for interpreting the breadth of the invention.
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|May 9, 1995
|Jan 25, 2000
|Curagen Corporation
|Apparatus and method for the generation, separation, detection, and recognition of biopolymer fragments
|Apr 26, 1999
|Apr 17, 2001
|Curagen Corporation
|Apparatus and method for the generation, separation, detection, and recognition of biopolymer fragments
|Apr 26, 1999
|May 22, 2001
|Curagen Corporation
|Apparatus and method for the generation, separation, detection, and recognition of biopolymer fragments
|Jun 21, 1999
|Nov 26, 2002
|Curagen Corporation
|Apparatus and method for the generation, separation, detection, and recognition of biopolymer fragments
|Apr 9, 2003
|Dec 26, 2006
|Tecan Trading Ag
|Cassette, system, and 2-D gel electrophoresis method for separating molecules
|Apr 23, 2003
|Sep 4, 2007
|Gene Bio-Application Ltd.
|Gel trap for electrophoresis
|Dec 4, 2002
|Dec 4, 2007
|Owl Separation Systems, Inc.
|Horizontal gel casting device and method
|Mar 8, 2006
|Jan 22, 2008
|Gene Bio-Application Ltd.
|Gel for electrophoresis
|Apr 23, 2003
|Feb 5, 2004
|Yitzhak Ben-Asouli
|Gel for electrophoresis
|Apr 23, 2003
|Feb 5, 2004
|Yitzhak Ben-Asouli
|Gel trap for electrophoresis
|Apr 9, 2003
|Mar 11, 2004
|Nikolaus Ingenhoven
|Cassette, system, and 2-D gel electrophoresis method for separating molecules
|Nov 7, 2003
|Sep 22, 2005
|Roger Patrick
|Methods and apparatus for optimizing a substrate in a plasma processing system
|Mar 8, 2006
|Sep 21, 2006
|Gene Bio-Application Ltd.
|Gel for electrophoresis
|Jan 5, 2007
|Jul 10, 2008
|Artz Matthew R
|Rapid gel electrophoresis system
|Apr 11, 2003
|Oct 15, 2003
|Tecan Trading AG
|Cassette, system, and 2D gel electrophoresis method for separating molecules
|Apr 26, 2000
|Nov 2, 2000
|Mj Research, Inc.
|Electrophoresis assembly and method of casting electrophoresis gels
|Oct 29, 2001
|May 10, 2002
|Gene Bio-Application Ltd.
|Gel trap for electrophoresis
|Oct 29, 2001
|Jul 25, 2002
|Gene Bio Applic Ltd
|Gel trap for electrophoresis
|Oct 29, 2001
|May 10, 2002
|Gene Bio-Application Ltd.
|Gel for electrophoresis
|Oct 29, 2001
|Jul 25, 2002
|Gene Bio Applic Ltd
|Gel for electrophoresis
|U.S. Classification
|International Classification
|Cooperative Classification
|European Classification
|Jun 28, 1994
|Certificate of correction
|Jan 9, 1997
|Fee payment
Year of fee payment: 4
|Dec 22, 2000
|Fee payment
Year of fee payment: 8
|Dec 17, 2004
|Fee payment
Year of fee payment: 12