Navigation Links
Easily Amplify Genomic DNA with Long-Distance PCR


Using TaqPlus Long DNA polymerase to genotype mice

Evgeny Loukianov Tanya Loukianova Muthu Periasamy
Cardiovascular Research Center, University of Cincinnati, Cincinnati, OH

We developed a simple, reliable procedure to genotype mice using long-distance PCR, whereby genomic DNA is isolated from mouse ear clips, and large DNA fragments are amplified using TaqPlus Long DNA polymerase.*,,

PCR is the method of choice for screening transgenic mice1 because it is fast, allows a large number of samples to be screened at one time, is less laborious than Southern blotting, and does not require purified DNA or the use of radioactive materials. Genomic DNA for PCR screening of mice is generally prepared from tail biopsies2 or blood samples3; however, the routine procedure of clipping the ear or toe to mark the mice4 provides enough tissue to extract DNA for PCR amplification. The ear clip procedure is advantageous because not only are mice identified and genotyped simultaneously, they tolerate ear clipping better than cutting of the tail or toes.

In many cases, it is beneficial to amplify large DNA fragments to verify the structure of a modified genetic locus. For example, when using targeting vectors with long arms,5 conventional PCR is not suitable for mouse genotyping.6 Hence, we developed a procedure to amplify large DNA fragments with TaqPlus Long DNA polymerase using genomic DNA from mouse ear clips.

Genomic DNA Prepared from Mouse Ear Clips

The integrity of the DNA template is crucial for reproducible long-distance PCR.7 Because DNA prepared from ear clips by standard methods2,8 failed to provide reproducible amplification of large (1.5 kb) DNA fragments, we developed a
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Printable CELLocate Log Sheets for Easily Locating Adhererent Cells
2. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
3. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
4. New Protocols for Isolating High- Molecular-Weight Genomic DNA
5. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
6. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
7. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
8. Mouse Tail Genomic DNA Isolation Protocol(1)
9. Genomic DNA Isolation Protocol(1,2,3)
10. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
11. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Easily Amplify Genomic DNA with Long Distance PCR

(Date:12/16/2009)... has what scientists have been asking for! Zap-CHO is a ... and productivity of CHO cells. Developed for scientists seeking maximum ... defined and consistent recombinant CHO supplement. , ... ... Scientists working with CHO cells can now access an informational ...
(Date:12/15/2009)... Department of Energy (DOE) bio-energy research centers, has selected ... intellectual property. , ... ... Surety, LLC , the leading provider of data ... BioEnergy Institute (JBEI), one of three Department of Energy ...
(Date:12/15/2009)... WILMINGTON, Del. and CHARLOTTESVILLE, Va., Dec. 15 ... ) and the University of Virginia (UVa) ... entered into a strategic research collaboration to ... coronary artery disease (CAD) with a secondary ... (Logo: http://www.newscom.com/cgi-bin/prnh/20091027/PH99766LOGO ) ,, The ...
(Date:12/15/2009)... N.Y., Dec. 15 On January 20th, ... Society for Medical Publication Professionals (ISMPP) will be ... London, UK. More than 100 attendees are expected ... for publication professionals, editors, publishers, medical writers and ... medical publications and the impact of guidelines and ...
Breaking Biology Technology:Online Video of InVitria's Zap-CHO Helps Scientists Maximize CHO Cell Culture Performance 2Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 2Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 3Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 4Joint BioEnergy Institute Selects Surety's AbsoluteProof to Protect the Integrity and Legally Defend the Authenticity of its Scientific Intellectual Property 5AstraZeneca and the University of Virginia Sign Strategic Research Collaboration to Develop Novel Treatments for Cardiovascular Disease 2AstraZeneca and the University of Virginia Sign Strategic Research Collaboration to Develop Novel Treatments for Cardiovascular Disease 3Announcing ISMPP EU 2010 - the First European Meeting of the International Society for Medical Publication Professionals, Royal College of Physicians, London, UK, January 20, 2010 2
... issue, Wisconsin Technology Network launches a special section devoted ... is not new, it is only now beginning to ... that perspective that we present this series of articles ... by Wisconsin experts in business and academia. , ,In ...
... -- Third Wave Technologies Inc. has filed suit ... two products, the company announced on Tuesday. The suits were ... Friday filed suit against Digene Corp., seeking a ... papilloma virus (HPV) products. , ,On Tuesday, Third Wave filed ...
... parents' generation of corporate leaders. , ,A quarter-century ago, ... lips of gray-haired, gray-suited corporate executives. Today, cool is ... yet have it. , ,At the Olympia Resort on ... arts groups met to brainstorm strategies for making Wisconsin ...
Other Biology Technology:RFID starting to make real impact on business 2RFID starting to make real impact on business 3Third Wave files patent suits against Digene, Chiron and Bayer 2Tech leaders call for more creativity 2
(Date:12/16/2009)... BIO-key International, Inc. (OTC Bulletin ... safety and finger-based biometric identification solutions, announced ... a Special Shareholder Meeting held today approved ... Division to Interact911 Mobile Systems Inc. Of ... favor and 1.2% were opposed. ,, (Logo: ...
(Date:12/16/2009)... WALL, N.J., Dec. 9 BIO-key ... leader in finger-based biometric identification solutions, today ... world,s most respected security publications, named BIO-key ... in the Access Control sub-category Biometrics. ,, ... to Peter Stephenson, technology editor, SC Magazine, ...
(Date:12/15/2009)... The 2009 Frost & Sullivan ... presented to Sagem Securite (Safran Group) for ... award recognises the company,s expertise in advanced ... offering novel, value-added products to end users. ... ,, "Sagem Securite has been involved in ...
Breaking Biology News(10 mins):BIO-key(R) Shareholders Overwhelmingly Approve Sale of Law Enforcement Division 2BIO-key(R) Shareholders Overwhelmingly Approve Sale of Law Enforcement Division 3BIO-key(R) Selected as 2009 Industry Innovator in Biometrics by SC Magazine 2BIO-key(R) Selected as 2009 Industry Innovator in Biometrics by SC Magazine 3Frost & Sullivan Recognises Sagem Securite for its Innovative Automated Border Control Systems 2Frost & Sullivan Recognises Sagem Securite for its Innovative Automated Border Control Systems 3Frost & Sullivan Recognises Sagem Securite for its Innovative Automated Border Control Systems 4IPC The Hospitalist Company Acquires Two New England Practices 61664 1IPC The Hospitalist Company Acquires Two New England Practices 61664 2Mylans President of Europe Middle East and Africa Elected President of European Generic Medicines Association 6393 1Mylans President of Europe Middle East and Africa Elected President of European Generic Medicines Association 6393 2MS Need Not Preclude Pregnancy 61660 1MS Need Not Preclude Pregnancy 61660 2
... University of Pennsylvania School of Medicine have discovered that ... used to study HIV in macaque monkeys provides the ... an unmodified vaccine. This multi-year study found that the ... effects of a vaccine derived from the DNA of ...
... portions of the human genome with those of other ... unidentified human genes, and found extensions of several hundred ... idea that as organisms evolve, sections of genetic code ... different ways., The research is reported by Adam Siepel, ...
... PITTSBURGH Carnegie Mellon University graduate engineering students Shahzeen Attari, ... Samaras are first-place winners in a letter-writing competition called ... ... by Johnson Controls Inc., asked students to demonstrate their ...
Other Biology News:Penn researchers find monkeys able to fend off AIDS-like symptoms with enhanced HIV vaccine 2Evolutionary comparison finds new human genes 2Evolutionary comparison finds new human genes 3Carnegie Mellon students win contest 2Carnegie Mellon students win contest 3
... unique human proteins on a 1x3 ... a cross section of all cellular ... high-quality results. Additional control proteins and ... validity and reproducibility., ,UNIchip protein microarrays ...
... UNIchip AV-400 contains 400 unique ... nitrocellulose-coated glass slide. The proteins, all ... Ontology classification, are arrayed in quadruplicate ... and a specific chip layout guarantee ...
...
... E.Coli glycerol stocks contained in standard 96-well ... clones with approximately 5,000 clones per plate. ... plate well, the corresponding subplate is screened ... the positive well(s). Cells from a positive ...
Biology Products: