We suggest adding 10 ng Tte UvrD Helicase to a 25 μl LAMP reaction using the WarmStart® LAMP Kit (NEB #E1700). Helicase should be added to all reactions, as it has minimal effect on performance of reactions containing the target DNA or RNA (though fluorescence signal may decrease; reaction time should not be affected). For reactions with very high non-template amplification levels, the amount of helicase may need to be increased. If positive signal is affected, the amount of helicase should be reduced. Suggested ranges to use in LAMP reactions with indicated DNA polymerases are as follows:
Bst DNA Polymerase, Large Fragment (NEB #M0275): | 5-10 ng per reaction (0.2–0.4 ng/μl) |
Bst 2.0® DNA Polymerase (NEB #M0537): | 10-15 ng (0.4–0.6 ng/μl) |
Bst 3.0 DNA Polymerase (NEB #M0374): | 10–20 ng (0.4–0.8 ng/μl) |