"
Purdue/3 July 2008
From 2008.igem.org
(Difference between revisions)
(New page: Click Here to return to the notebook.) |
|||
| Line 1: | Line 1: | ||
[[Team:Purdue/Notebook | Click Here to return to the notebook.]] | [[Team:Purdue/Notebook | 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''' | ||
Latest revision as of 17:07, 3 July 2008
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
