Team:University of Ottawa/25 June 2008

From 2008.igem.org

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:'''PCR amplification of S, D, T, 7 and 8'''
:'''PCR amplification of S, D, T, 7 and 8'''
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::<li> S, D, and T plasmids were PCR amplified using primers F69 and F70
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::<li> S, D, and T plasmids were PCR amplified overnight using primers F69 and F70
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::<li> PCR program for 0&1 was: PHUSION
 +
::<li> PCR program for S&D&T was: PHN1
::<li> 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 )
::<li> 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 )
:::{| class="wikitable"
:::{| class="wikitable"

Revision as of 17:56, 30 June 2008

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