Bench work
- PCR cleanup
- 70μL of membrane binding solution added directly to BSA PCR tube, for a final volume of 140μL (some of the extra volume is from the wax, but this is still more than expected assuming that I used 15μL from this tube last week).
- followed manufacturer protocol, using the vacuum method, and eluted with 50μL H2O
- NheI digest
- 15μL of cleaned-up BSA PCR from step 1 + 5μL NEB buffer 2 + 24.5μL H2O + 0.5μL 100X BSA + 5μL NheI
- → 2h@37°C
- → 20'@65°C
- → store @ -20°C
|