Purdue/3 July 2008

From 2008.igem.org

Revision as of 17:07, 3 July 2008 by Mstine (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Click Here to return to the notebook.

Transformation!!!

Today we're finishing the transformation we planned earlier this week. The parts to be transformed are:

  • J09855:
    • Constitutive luxR device w/ pLuxR
    • Plate 1003, Well 5B
    • QC Confirmed/OK
    • AmpR, psB1A2
    • This has been prepped and soaking in TE at room temp. for 2 days
  • J13003 (Using 2007 DNA):
    • POPS + RIPS generates CFP. POPS generates YFP.
    • Plate 2, Well 3H
    • We will add 15uL of diH2O to prep it, and add 1uL of the mixture for transformation

Procedure

  • Place DNA samples on ice
  • Add 5uL DNA/TE solution or 1uL DNA/water solution to 25uL of competent cells.
  • Tap very gently to mix
  • Incubate cells + DNA on ice for exactly 30 minutes
  • Incubate EXACTLY 30 seconds in 42C water bath. DO NOT MIX OR SHAKE
  • Remove vials--DO NOT MIX OR SHAKE
  • Add 250uL of SOC (pre-warmed)
  • Shake vails at 37C for exactly 1 hour at 225 rpm
  • Spread 100uL on amp plates, incubate at 37C
  • Put remaining solution at 4C and store for later


Edited by Janie Stine

Retrieved from "http://2008.igem.org/Purdue/3_July_2008"