Description
The Uracil-Specific Excision Reagent (referred to as USER) is an enzyme cocktail consisting of Uracil DNA Glycosylase (UDG) and Endonuclease VIII (Endo VIII). It is capable of generating single-nucleotide gaps specifically at uracil locations. UDG facilitates the cleavage of the uracil base, resulting in the formation of an AP site (apurinic/apyrimidinic site) while maintaining the integrity of the phosphodiester sugar-phosphate backbone. The endonuclease activity of Endo VIII cleaves the phosphodiester bonds at both the 3′ and 5′ ends of the apurinic site, thereby releasing the deoxyribose sugar that is lacking a base.
Features
No residual nucleases or RNases
High excision efficiency
Applications
RNA strand-specific library construction
Uracil-specific excision-mediated cloning (USER-LIC)
DNA assembly, such as the assembly of TALE monomers
Serial construction of multiple cDNA libraries from single cells
Specifications
|
Concentration |
1 U/µL |
|
Unit Definition |
One Unit is defined as the amount of enzyme required to nick a 34-mer double-stranded oligonucleotide containing a single uracil base in a 10 μL reaction system at 37°C for 15 minutes, which corresponds to the cleavage of 10 pmol of the substrate. |
|
Inactivation Conditions |
75℃, 10 min |
Components
|
Component No. |
Name |
14537ES50 |
14537ES72 |
|
14537-A |
Uracil-Specific Excision Reagent (1 U/μL) |
50 μL |
250 μL |
|
14537-B |
10×Uracil-Specific Excision Reagent Reaction Buffer |
1.25 mL |
1.25 mL |
Shipping and Storage
The products should be stored at -25℃ ~ -15℃ for 1 year.
Figures
Figure1. Comparison of excision effects
Note: 10 pmol of double-stranded DNA containing dUTP was excised using Yeasen's USER enzyme and a comparable product from Supplier A. The results from agarose gel electrophoresis showed that the excision effects of Yeasen's USER enzyme were consistent with those of Supplier A
Figure2. Comparison of colony counts for USER-LIC cloning
Note: A 600 bp DNA fragment was ligated using Yeasen's USER enzyme and a comparable product from Supplier A to construct a vector, which was then transformed into E. coli and cultured. The number of colonies on the culture plate was observed, and the results indicated that the USER-LIC cloning efficiency using Yeasen's USER enzyme was superior to that of Supplier A.
Documents:
Safety Data Sheet
Manuals
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