Protocol for Exonuclease VII (M0379)

Protocol for removing primers with 3' modifications from a PCR reaction

1. Set-up the reaction as follows:

Reaction Volumes
 PCR Product
5 μl
 Exonuclease VII
 2 μl (20 units)

2. Incubate the mix at 37°C for 30 minutes.

3. Heat inactivation 95°C for 10 minutes prior to Sanger DNA sequencing. For other downstream molecular cloning applications column cleanup is recommended.

4. To clean up treated samples, we recommend using one of the following steps:

a. Column clean up (we recommend the Monarch® PCR & DNA Cleanup Kit, NEB #T1030), or

b. Running the reaction on an agarose gel, and then extracting the DNA (we recommend the Monarch Gel Extraction Kit, NEB #T1020), or

c. Performing a phenol/chloroform extraction followed by ethanol precipitation.

Note: See Exo-CIP™ Rapid PCR Cleanup Kit (NEB #E1050) for Sanger Sequencing, SNP detection, or library preparation for NGS.