Purdue/3 July 2008

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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