biotechrabbit™ A-Tailing Kit (Cat No. BR1101701) contains the essential reagents required for the addition of an adenine nucleotide to the 3' ends of DNA fragments. This kit includes a 20X A-Tailing Mix, a 1 mM dATP solution, and a 10X Activation Buffer. The reaction is designed to be performed in a single step and ensures a highly efficient A-tailing process, with minimal risk of overhang loss. The kit components are optimized to work together, providing consistent results with high specificity.
Component | Composition |
20X A-Tailing Mix | Optimized 20X A-Tailing Mix contains Taq DNA polymerase |
1 mM dATP | Aqueous solution (pH 7.0) of 1 mM 2'-deoxyadenosine 5'-triphosphate sodium salt |
10X Activation Buffer | Optimized A-Tailing buffer in 10X concentration |
STORAGE | Store at −20 °C until expiry date. |
Quality Control
Functional Assay
Product was functionally tested in the adenylation reaction of a blunt-ended DNA fragment. Presence of the A-tailed DNA fragment was confirmed using TA Cloning, followed by colony PCR.
REACTION SETUP
Prepare the reaction mix according to the table below.
Component |
Volume |
Final concentration |
DNA sample |
x µl |
100–500 ng |
20X A-Tailing Mix |
2.5 µl |
1X |
1 mM dATP |
10 µl |
0.2 mM |
10X Activation Buffer |
5 µl |
1X |
Nuclease-free water |
to 50 µl |
|
Incubation
- Incubate the mixture at 72 °C for 15–30 minutes.
- Keep the above DNA fragment with additional A residue at 3' end at −20 °C for later use.
Downstream Application
- The A-tailed DNA is now ready for use in cloning, sequencing, or NGS library preparation.
- Proceed with your downstream applications according to the relevant protocols.
Storage of Reaction Products
- If not used immediately, store the reaction product at −20 °C.
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