Home FAQs What is the best reaction volume for removal of linear dsDNA?

FAQ: What is the best reaction volume for removal of linear dsDNA?

We recommend incubating a 30 µl reaction containing 10 units of Exonuclease V for 30 minutes at 37°C in order to remove 1 µg of linear dsDNA ranging from 100-1500 bp. One can estimate the amount of linear DNA present by agarose gel electrophoresis or nanodrop measurement. Treated DNA can be further purified by ethanol precipitation or spin-columns (e.g., Monarch PCR & DNA Cleanup Kit (NEB T1030).

Recommended reaction setup (For removal of less than 1 µg/ul DNA):
 
dsDNA x μl
10 x NEBuffer 4 3 µl
10 mM ATP 3 µl
H2O 23-x µl
Exonuclease V
1 µl
Total 30 µl