Team:UCLA/Notebook/Spider Silk Genetics/28 July 2015

iGEM UCLA




7/28/2015

Miniprep and Sequencing

  • Miniprep (qiagen) for M2-12(1C3) liquid culture.
  • Sequencing with VF, VR.

Maxiprep for M2-AB, BC, CA

  • Maxiprep using qiagen kit.
    • M1-AB: 111 ng/uL
    • M1-BC: 150 ng/uL
    • M1-CA: 350 ng/uL

BsaI Digestion for M1-AB, BC, CA

  • Olivia digested 5 ug of relevant plasmid in a 50 uL reaction with 4 uL of BsaI.
    • Digest 50 C 2 hrs, heat kill 65 C 20 min.
  • Run on 1.5% TAE gel to visualize. Used 3 uL of NEB 100 bp ladder.
Fig. 1 BsaI Digestion of M1-AB, BC, CA. The expected product size is 102 bp.
  • Excised the 102 bp bands for gel extraction
  • Also saved some of the digested backbone as linearized pSB1C3.