Navigation Links
Uracil-DNA Glycosylase

for prevention of carry-over contamination in PCR Cat. No. 1 444 646 100 units
Cat. No. 1 269 062 25 units Description Uracil-DNA Glycosylase (UNG) (from E. coli) catalyzes the removal of uracil from single- and double-stranded DNA that has been synthesized in the presence of dUTP. The apyriminic sites formed by UNG are susceptible to cleavage by heat under alkaline conditions (carry-over prevention). UNG will not digest 3'-terminal dU, dUTP, primers labeled with biotin-dUTP, or digoxigenin-dUTP. Ribouracil residues in RNA are also unaffected by UNG. Application Prevention of carry-over contamination in:
  • PCR with Taq DNA Polymerase.
  • RT-PCR with C. therm. Polymerase One-step RT-PCR System.
Operating parameters
  • Substrate: Single- or double-stranded DNA containing deoxyuracil (DNA synthesized in the presence of dUTP).
  • Incubation conditions for cleavage of uracil-glycosidic bonds: 20C, 110 min.
  • Thermal stability (pH 8.3): half-life at 40C, 2 min; half-life at 45C, 0.5 min.
  • MgCl2 concentration: 2.5 mM.
  • dNTP concentration: 200 M dNTPs; 600 M dUTP.
  • Conditions for inactivation of enzyme: 95C, 2 min.

Notes:

UNG (2 units) will degrade up to 106 uracil-containing DNA molecules in 10 min. In contr ast to the enzyme from E. coli, heat-labile UNG will not modify dU-containing PCR products for at least several hours at 4C. Therefore, it is not necessary to freeze dU-containing PCR products immediately after amplification, or to hold the reaction mixture at 72C until it is used. However, note that after prolonged storage (>8 h) residual enzyme activity may lead to degradation of PCR product. For long term storage, PCR product should therefore be frozen at 20C.

Key advantages
  • Prevents carry-over contamination in PCR, because the enzyme modifies all uracil-containing products from previous PCRs. These products are then hydrolyzed by heat under alkaline conditions.
  • Does not degrade native (thymidine-containing) templates, because the enzyme is free of contaminating exo- and endonucleases.
  • Does not interfere with PCR amplification, because the enzyme is not active at temperatures typically used for PCR applications.

'"/>

Source:


Page: All 1 2

Related biology technology :

1. Preventing Carry-over Contamination with Uracil-DNA Glycosylase
2. Uracil-DNA Glycosylase, heat-labile
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/27/2016)... June 27, 2016  Sequenom, Inc. (NASDAQ: ... healthier lives through the development of innovative products and ... the United States denied its petition ... claims of Sequenom,s U.S. Patent No. 6,258,540 (",540 Patent") ... established by the Supreme Court,s Mayo Collaborative Services v. ...
(Date:6/27/2016)...  Liquid Biotech USA , ... Sponsored Research Agreement with The University of Pennsylvania ... cancer patients.  The funding will be used to ... clinical outcomes in cancer patients undergoing a variety ... employed to support the design of a therapeutic, ...
(Date:6/24/2016)... ... 24, 2016 , ... Researchers at the Universita Politecnica delle Marche in Ancona ... or pleural mesothelioma. Their findings are the subject of a new article on the ... are signposts in the blood, lung fluid or tissue of mesothelioma patients that can ...
(Date:6/23/2016)... A person commits a crime, and the detective ... the criminal down. An outbreak of foodborne illness ... (FDA) uses DNA evidence to track down the bacteria that ... It,s not. The FDA has increasingly used a complex, cutting-edge ... illnesses. Put as simply as possible, whole genome sequencing is ...
Breaking Biology Technology:
(Date:6/22/2016)... , June 22, 2016   Acuant ... and verification solutions, has partnered with RightCrowd ... solutions for Visitor Management, Self-Service Kiosks and ... products that add functional enhancements to existing ... corporations and venues with an automated ID ...
(Date:6/16/2016)... 16, 2016 The global ... to reach USD 1.83 billion by 2024, according ... Inc. Technological proliferation and increasing demand in commercial ... to drive the market growth.      ... The development of advanced multimodal techniques for biometric ...
(Date:6/7/2016)...  Syngrafii Inc. and San Antonio Credit Union ... integrating Syngrafii,s patented LongPen™ eSignature "Wet" solution into ... result in greater convenience for SACU members and ... existing document workflow and compliance requirements. ... Highlights: ...
Breaking Biology News(10 mins):