BioBrick Transformations
Plasmid Isolation
PCRs: Adding Restriction Sites and Linkers
Restriction Digests
Restriction enzymes used in our assembly | |||||||
---|---|---|---|---|---|---|---|
Restriction Enzyme | Sequence | Activity in NEBuffers (%) | Incubation temperature | Heat inactivation | |||
1.1 | 2.1 | 3.1 | CutSmart | ||||
AgeI | A\CCGGT | 100 | 75 | 25 | 75 | 37°C | 65°C |
AgeI-HF | A\CCGGT | 100 | 50 | 10 | 100 | 37°C | 65°C |
BamHI | G\GATCC | 75* | 100* | 100 | 100* | 37°C | — |
BamHI-HF | G\GATCC | 100 | 50 | 10 | 100 | 37°C | — |
HindIII | A\AGCTT | 25 | 100 | 50 | 50 | 37°C | 80°C |
HindIII-HF | A\AGCTT | 10 | 100 | 10 | 100 | 37°C | 80°C |
HindIII | A\AGCTT | 25 | 100 | 50 | 50 | 37°C | 80°C |
HindIII-HF | A\AGCTT | 10 | 100 | 10 | 100 | 37°C | 80°C |
NcoI | C\CATGG | 100 | 100 | 100 | 100 | 37°C | 80°C |
NcoI-HF | C\CATGG | 50 | 100 | 10 | 100 | 37°C | 80°C |
NheI | G\CTAGC | 100 | 100 | 10 | 100 | 37°C | 65°C |
NheI-HF | G\CTAGC | 100 | 25 | 10 | 100 | 37°C | 80°C |
* denotes star activity| NOTE ADD NdeI TOO |