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:10/11/2017)... ... October 11, 2017 , ... Proscia Inc ., a ... Webinar titled, “Pathology is going digital. Is your lab ready?” with Dr. Nicolas ... practices and how Proscia improves lab economics and realizes an increase in diagnostic ...
(Date:10/11/2017)... Florida (PRWEB) , ... October 11, 2017 , ... ... Drug Administration (FDA) has granted orphan drug designation to SBT-100, its novel anti-STAT3 ... for the treatment of osteosarcoma. SBT-100 is able to cross the cell membrane ...
(Date:10/10/2017)... , Oct. 10, 2017 International research firm Parks Associates ... will speak at the TMA 2017 Annual Meeting , October 11 ... in the residential home security market and how smart safety and security ... Parks Associates: ... "The residential security ...
(Date:10/9/2017)... ... 09, 2017 , ... The award-winning American Farmer television series will feature 3 ... airs Tuesdays at 8:30aET on RFD-TV. , With global population estimates nearing ten ... to continue to feed a growing nation. At the same time, many of our ...
Breaking Biology Technology:
(Date:4/5/2017)... April 5, 2017  The Allen Institute for Cell ... Explorer: a one-of-a-kind portal and dynamic digital window into ... data, the first application of deep learning to create ... cell lines and a growing suite of powerful tools. ... these and future publicly available resources created and shared ...
(Date:4/4/2017)... , April 4, 2017   EyeLock LLC , ... that the United States Patent and Trademark Office (USPTO) ... covers the linking of an iris image with a ... and represents the company,s 45 th issued patent. ... is very timely given the multi-modal biometric capabilities that ...
(Date:3/30/2017)... The research team of The Hong Kong ... identification by adopting ground breaking 3D fingerprint minutiae recovery and matching ... and accuracy for use in identification, crime investigation, immigration control, security ... ... A research team led by ...
Breaking Biology News(10 mins):