Boston UniversityRestrictionEnzymePlasmid
From 2007.igem.org
Plasmid and Restriction Enzyme Choice
Using pJQ200 plasmid.
sacB site
Enzyme Cat# Temp Supplied NEBuffer Supplements % Activity in NEBuffer BSA SAM 1 2 3 4 HindIII R0104 37°C NEBuffer 2 No No 50 100 10 50 EcoRI R0101 37°C NEBuffer EcoRI No No 100 100 100 100
Double Digest Recommendation(s) for HindIII + EcoRI: Digest in NEBuffer EcoRI at 37°C. Note: The above recommendation is based on the experimental results. Please check Suggested NEBuffers for Double Digestion.
gmR site
Enzyme Cat# Temp Supplied NEBuffer Supplements % Activity in NEBuffer BSA SAM 1 2 3 4 BsaI R0535 50°C NEBuffer 3 No No 75 75 100 50 Tth111I R0185 65°C NEBuffer 4 No No 50 25 25 100
Double Digest Recommendation(s) for BsaI + Tth111I: Digest in NEBuffer 4 at 50°C with BsaI, then add Tth111I and raise temperature to 65°C. At least one enzyme has < 100% activity in this buffer, so additional units of enzyme and/or longer incubation time may be necessary.