"
Team:University of Ottawa/25 June 2008
From 2008.igem.org
Contents |
Today in the Lab
Matt
- PCR Amplification
- F60, F61 primers used for PTP2 cassette. I also labelled all the primers we received that we will be using for the next few months.
- I decided to use the Kaern Lab protocol for PCR Amplification.
- The PTP2 PCR product was then run on a gel - gel results were as expected with 2275 size band.
- Glycerol Stock
- Glycerol stock of plasmids D12, S1, T123, pSS042 with LB + glycerol was performed.
Dan
- Ran gel of pSSA42, S1, D12, T123 plasmids
- This time the marker was added and expected results were obtained. SB, DA, TB, and 42B colonies were chosen for glycerol stock.
- Cleaned out DNA box
- Cleaned out all DNA that will not be used from our box
- Our names are getting complicated so I am renaming our DNA using the following legend (all other DNA was discarded)
Plasmid Strain New Name 587 7B 7 598 8B 8 600 0A 0 601 1C 1 S1 SB S D12 DA D T123 TB T pSSA42 42B 42
- PCR amplification of S, D, T, 7 and 8
- S, D, and T plasmids were PCR amplified overnight using primers F69 and F70
- PCR program for 0&1 was: PHUSION
- PCR program for S&D&T was: PHN1
- The 0 and 1 plasmids were amplified according to the following chart: (Primer F66 is :Int TetR Repl F (*PstI) (B). primer F67 is Int GFP Repl F(*SphI) (A), and primer F68 is:Int SSRE R(*ClaI )
Sample F66 F67 F68 0A Y Y 1A Y Y 0B Y Y 1B Y Y
