Korea U Seoul/1 October 2010
From 2010.igem.org
Digestion
- Check : recombinant plasmid sequence
- Selection of correct clones
- Digestion of PCR products(ParsR-GFP) and pSB1C3
- ParsR-GFP : EcoRI and SpeI
- pSB1C3 : EcoRI and PstI
Reagent | Volume (μL) |
DNA (about 30ng/μL) | 30 |
10x NEB buffer 2 | 5 |
BSA (10mg/mL) | 0.5 |
Appropriate 1st and 2nd restriction enzymes | 2 (each 1) |
D.W. | 12.5 / total = 50 |
Completely digestion at 37˚C for 2 hours (at least)
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- Confirmation of digested products by agarose gel electrophoresis (Figure 6)
- Quantification of DNA concentration by NanoDrop