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  1. Run Gel on digested E1010 and diagnostic ladder
  2. PCR Purify parts
  3. Finish standard assembly of vector + E1010 insert
  4. Check sequencing results
  5. Make Amp+Kan plates and do a colony screening

Running Gels

  • 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 10┬ÁL (Amp diluted 10X, will be different number for adding Kan to Amp) per mL of LB+Kan poured on plate.
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