Boston UniversityRestrictionEnzymePlasmid

From 2007.igem.org

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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.