Protocol for PCR Product Clean-up

Protocol for PCR Product Clean-up

(with Exonuclease I , E.coli and Calf Intestine Alkaline Phosphatase
or Shrimp Alkaline Phosphatase)

1. Prepare the following reaction mixture:

PCR mixture (directly after completion of PCR) 5µl

Exonuclease I
0.5µl(10u)

Calf Intestine Alkaline Phosphatase 1µl
or Shrimp Alkaline Phosphatase

2. Mix and incubate at 37 degree for 15min.

3. Stop the reaction by heating the mixture at 85 degree for 15 min.

Note:

Up to 5µl of the purified PCR products can be used directly for DNA
sequencing without further purification.

the protocol may be applied for clean-up of PCR products, generated by
any thermophilic DNA polymerase or polymerase mix.

the procedure is not recommended for downstream cloing application.