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:5/27/2015)... , May 27, 2015  Ceres, Inc. (Nasdaq: ... a U.S. patent for a genetic sequence derived from ... as research, product development and plant transformation. The company ... biosynthetic processes that are the target of a class ... companies a commercial license to the innovation, including an ...
(Date:5/26/2015)... and BELLINGHAM, Washington, USA (PRWEB) May 26, 2015 ... signed a formal partnership with the Max Planck ... for Extreme and Quantum Photonics. The center will link ... field of photonics, and will be only the third ... of SPIE, the international society for optics and ...
(Date:5/26/2015)... California and HILDEN, Germany ... QIAGEN N.V. (NASDAQ: QGEN ; Frankfurt ... of its QIAGEN Clinical Insight ® (QCI™) ... labs to interpret and report on genomic variants ... supported applications for the bioinformatics platform are in ...
(Date:5/26/2015)... FL (PRWEB) May 26, 2015 ... Inc. have announced plans for the company’s first cellular ... 2015, followed by a full day, hands-on stem cell ... by J. Victor Garcia Gimenez, M.D., President of Therapeutic ... the medical community’s increasing interest in regenerative medicine and ...
Breaking Biology Technology:Ceres Awarded Patent for Innovation in Soybean 2Ceres Awarded Patent for Innovation in Soybean 3Third Max Planck Centre in North America Inaugurated at University of Ottawa 2Third Max Planck Centre in North America Inaugurated at University of Ottawa 3QIAGEN Launches Clinical Bioinformatics Platform for Next-Generation Sequencing 2QIAGEN Launches Clinical Bioinformatics Platform for Next-Generation Sequencing 3QIAGEN Launches Clinical Bioinformatics Platform for Next-Generation Sequencing 4QIAGEN Launches Clinical Bioinformatics Platform for Next-Generation Sequencing 5Stem Cell Training, Inc. Announces First European Symposium on Cellular and Regenerative Medicine 2Stem Cell Training, Inc. Announces First European Symposium on Cellular and Regenerative Medicine 3
... Antonio Breast Cancer Symposium, AMSTERDAM, Netherlands, Dec. ... evolving field of molecular diagnostics, announced,today that an ... its MammaPrint(R) breast cancer prognosis test in,patients who ... that,MammaPrint(R) can accurately identify a low risk group ...
... Patients treated with drug-eluting,stents must take a ... at least one year, and possibly longer, after ... guideline update published,online today in the journals of ... College of Cardiology (ACC), and,American Heart Association (AHA). ...
... Dec. 13 Sangart, Inc., a privately held,biopharmaceutical ... oxygen transport agents, today announced that an,independent data ... both Phase III clinical trials of Sangart,s lead ... recommendation follows the DSMB,s review of the blinded ...
Cached Biology Technology:MammaPrint(R) Breast Cancer Test Validated in Lymph Node-Positive Breast Cancer Patients 2MammaPrint(R) Breast Cancer Test Validated in Lymph Node-Positive Breast Cancer Patients 3PCI Guideline Update Stresses Long-Term Anticoagulant Therapy After Treatment With Drug-Eluting Stents 2PCI Guideline Update Stresses Long-Term Anticoagulant Therapy After Treatment With Drug-Eluting Stents 3PCI Guideline Update Stresses Long-Term Anticoagulant Therapy After Treatment With Drug-Eluting Stents 4Independent Data Safety Monitoring Board Recommends Continuation of Sangart's Pivotal Phase III Trials of Hemospan(R) 2
(Date:5/26/2015)... May 26, 2015 Research ... addition of the "Saudi Arabia Biometric Systems ... their offering. ,     (Logo: http://photos.prnewswire.com/prnh/20130307/600769) , ,The biometric ... is projected to grow at over 22% ... is attributed to the surging demand for better ...
(Date:5/21/2015)... May 21, 2015 According ... Market by Solutions (Hardware, Software, Services), by Applications (Surveillance ... others), by End-Users (Military & Defense & Commercial) ... Thermal Imaging Market is expected to grow from ... 2020, at a Compound Annual Growth Rate (CAGR) ...
(Date:5/19/2015)... -- Fingerprint Cards (FPC) has received an order ... FPC1035 from one if its module partners. Deliveries are planned ... and the sensors will be used by smartphone manufacturers in ... hereby amount to 740 MSEK to date. In addition to ... a number of smaller orders not separately communicated amounting to ...
Breaking Biology News(10 mins):Saudi Arabia Biometric Systems Market Forecast and Opportunities, 2020 with Morpho Safran, ZK Technlogy, and Suprema Dominating 2Thermal Imaging Market Worth $9,998.9 Million by 2020 2Thermal Imaging Market Worth $9,998.9 Million by 2020 3Thermal Imaging Market Worth $9,998.9 Million by 2020 4Fingerprint Cards Receives Touch Fingerprint Sensor Order of SEK 230 Million and Raises Revenue Guidance for 2015 2
... gum disease, is an independent risk factor for head ... for increased efforts to prevent and treat periodontitis as ... form of cancer. "Prevent periodontitis; if you have ... said Mine Tezal, D.D.S., Ph.D., assistant professor in the ...
... are developing diversionary tactics to fool disease-causing bacteria in ... gut infections, such as cholera, produce toxins that damage ... displayed on the surface of cells in the host,s ... meeting at Heriot-Watt University, Edinburgh, today (8 September), Professor ...
... reveal not only the "lifestyle" of marine microbes but of ... researchers may be able to predict the types of microbes ... of just a few dominant species, according to research co-author ... well, it may reveal new insights into the impacts of ...
Cached Biology News:Prevent periodontitis to reduce the risk of head and neck cancer 2Designing probiotics that ambush gut pathogens 2Genomes reveal bacterial lifestyles: Research 2
... designed for RNA aptamer screening and binding ... powerful Paraflo microfluidic on-chip synthesis platform. Thousands ... synthesized on-chip and LC Sciences can provide ... are available as part of our comprehensive ...
... is commonly used as a fusion partner ... The GSTTag sequence has been reported to ... the solubility of its fusion partners. When ... GSTTag fusion proteins can be purified with ...
... next step in the evolution of laboratory ... that delivers labware to multiple laboratory instruments, ... the throughput of ordinary microplate robots. It ... of configurations to meet any labs specific ...
... use of small interfering RNA (siRNA) molecules ... interference (RNAi) pathway have generated tremendous interest ... siRNA for experimental use can be synthesized ... then transfected into the target cells in ...
Biology Products: