rE. coli MutY with Buffer
rE. coli MutY with Buffer Summary
E. coli MutY DNA Glycosylase acts together with E. coli Formamidopyrimidine-DNA Glycosylase, or Fpg, to prevent the potentially mutagenic consequences of 8-oxo-dG lesions. 8-oxo-dG lesions escaping repair by Fpg frequently pair with Adenine (A) during DNA replication, producing an 8-oxo-dG:A mispair. MutY removes the A from this base pair to initiate base excision repair. In the absence of MutY, DNA replication after an 8-oxo-dG:A mismatch results in thymine incorporation opposite the adenine in one of the daughter strands, creating a fixed mutation. MutY has an associated AP lyase activity.
Assay Conditions and Analysis
4 pmoles of A/G mismatch oligonucleotide set with the A oligo end-labeled, 1X REC Buffer 4 (10 mM HEPES-KOH (pH 7.4), 100 mM KCl, 10 mM EDTA), and serial dilutions of enzyme in a 20 μL reaction volume are incubated for 1 hour at 37 °C. To complete cleavage of a basic site, fresh NaOH is added to final concentration of 166 mM then heated for 15 minutes at 95 °C. For analysis, 24 μL of 2X Loading Buffer (20 mM EDTA, 97% formamide, and 0.2% bromophenol blue) is added, the samples are heated at 95 °C for 5 min then fast cooled to 2-8 °C, and the cleavage products are resolved by 20% denaturing polyacrylamide gel electrophoresis. The bands are analyzed to quantify the cleavage products.
• 10X REC™ Buffer 4
• E. coli Mut Y DNA Glycosylase
- 10X REC™ Buffer 4
- E. coli Mut Y DNA Glycosylase
For research use only. Not for diagnositic use.
No product specific FAQs exist for this product, however you mayView all FAQs
Reviews for rE. coli MutY with Buffer
There are currently no reviews for this product. Be the first to review rE. coli MutY with Buffer and earn rewards!
Have you used rE. coli MutY with Buffer?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image