Team:Duke/Notebook/MayJun

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Revision as of 16:06, 29 July 2014

May 29

Objective: Prepare Chemically competent E. Coli
Matthew Faw

Followed Charlie’s Cloning Protocols to prepare chemically competent E. Coli

  • Once E. Coli were prepared, the solution was aliquoted into 12 labeled tubes (450l in each tube) and tubes were stored in iGEM box in -80C cooler on far left

Next Steps:

  • Transformation

June 1

Objective: Transform Chemically competent E. Coli (CCEC)
Matthew Faw

Transformation of CCEC with RFP Construct of varying concentrations+control with no DNA

  • Followed Charlie’s Cloning Protocols
    • Mistakenly used all of DNA construct from the kit… So not sure if the transformation will be successful
  • Plated the transformed DNA and put in incubator at 37C
  • Results (6/2/14)--Transformation unsuccessful because improper procedure was done by MFaw

Next Steps:

  • Look at plates, compare cultures

June 2

Objective: Redo yesterday’s transformation
Matthew Faw

Due to my failure to correctly conduct the transformation yesterday, it was necessary to redo the transformation.

  • Followed Charlie’s Cloning Protocols
    • Added 1 ul of .5, 5, 10, 20, 50, and 0 (control) pg/lof RFP Construct to chemically competent E. Coli (CCEC), and followed procedure
  • Plated the transformed DNA on 6 separate plates, put in 37C overnight
  • Results: (6/3/14)-Transformation unsuccessful. Try the transformation again with Charlie’s cells and with new cells grown using iGEM’s protocol