April 14th, 2010
- Prepare and grow strains with the required plasmids:
Strains were grown in LB medium with the correspondent antibiotic for each plasmid.
- Extract plasmids with Roche kit.
- Double resrtiction to extracted plasmids with EcoRI and Pst1.
Double restriction methods:
- DNA -> 5ul
- Buffer 2 -> 2ul (10% of total volume)
- BSA (required by PstI) -> 1ul
- EcoRI -> 1ul
- PstI -> 1ul
- HPLC -> 10ul (to complete total volume of 20ul)
- *Incubate at 37ºC overnight