- Run Gel on digested E1010 and diagnostic ladder
- PCR Purify parts
- Finish standard assembly of vector + E1010 insert
- Check sequencing results
- Make Amp+Kan plates and do a colony screening
- 85V for 45 Min
- E1010 digests came out fine
- Diagnostic Digest (used EcoRI and PstI to cut out insert, evaluated insert length) showed 3 bands.
- Eventually decided that the three bands were: desired vector, desired insert, F2620/B0034 vector.
- Will run colony screening using multi-antibiotic plates.
Making Amp+Kan plates
- If you want to add Amp resistance to Kan plates (or vice versa), add 10µL (Amp diluted 10X, will be different number for adding Kan to Amp) per mL of LB+Kan poured on plate.