Flat end DNA+A kit
Lot # Check on the product label
Size: 50 T
Storage: Store at -20°C.
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Price: $40.00
Catalog# BPC1098
Lot # Check on the product label
Size: 50 T
Storage: Store at -20°C.
Application: The PCR product amplified by a high-fidelity DNA polymerase with 3'→5' exonuclease activity such as Pfu, Pwo, Vent or KOD is a blunt-ended DNA fragment without A tail. If the blunt-ended DNA fragment needs to be combined with T When the vector is connected, it is necessary to use Taq DNA polymerase to add an A tail after the blunt-end fragment before it can be complementary to the T head of the T vector. This kit provides all the ingredients needed for A-tailing and can complete the entire process in about 20 minutes.
Components
Components |
Size |
Taq DNA polymerase |
50 µl |
10 x A-tailing buff er |
50 µl |
dATP Mix |
50 µl |
Protocol
1. The blunt-ended PCR product is cleaned by PCR (without non-specific amplification) or purified by gel recovery (with non-specific amplification).
2. The purified blunt-ended DNA fragments are A-tailed according to the following reaction system:
Blunt-ended DNA fragments |
7µl (up to 1µg) |
dATP Mix |
1µl |
10 x A-tailing buffer |
1µl |
Taq DNA polymerase |
1µl |
3. Mix gently and incubate at 72°C for 20-30 minutes.
4. The A-tailed product can be directly used in the ligation reaction without further cleaning or gel recovery and purification.