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Purdue/3 July 2008
From 2008.igem.org
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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
