Team:UCLA/Notebook/Spider Silk Genetics/31 August 2015

iGEM UCLA




8/31/2015

Transformation Results

  • All plates have colonies!

Starter Cultures Analysis

  • Starter cultures grown at 30 C are overgrown.
  • Cultures at 20 C show nothing.

Miniprep for M1-SeqAB2

  • Used qiagen kit to prep all 11 mL of culture.
  • 419.90 ng/uL A: 1.88

BsaI Digestion for M1-SeqAB2

  • Digest in 2x 50 uL reactions 5 ug of plasmid using 4 uL of BsaI.
  • Digest at 50 C for 2 hrs, heat kill 65 C for 20 min.
  • Visualize on 1% TAE gel, with 2 uL of NEB 50 bp ladder.
Fig. 1 BsaI digestion of M1-SeqAB2. The expected size is 102 bp.
  • Excised the 102 bp band, and extracted.
    • ~25 ng/uL

Liquid Culture

  • Picked 3 colonies from each plate: M1-12(1C3)/(T7), M1/2[1:2]-12(1C3)/(T7) into 5 mL liquid culture with chloramphenicol.
  • Grew overnight at 37 C.