Team:The University of Alberta/7 August 2008

From 2008.igem.org

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**Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands.
**Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands.
***Ligated the gel purified product with J61003.
***Ligated the gel purified product with J61003.
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'''Chris'''
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*PCR'd Tryp out of pUC57 (the original that Mike supplied)...buuut I used Vf and Vr to do it. I shouldnt have gotten any results because pUC57 doesnt have sites for those primers to bind BUT I did get a band about the right size...nevertheless, whatever was amplified wouldnt have the biobrick suffix and prefix in it so it was discarded.
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*Redid the PCR with the correct primers....got a good, sharp band ~1500bp in size BUT the same band was in the negative control. I didnt want to risk using something that was just a random 1500bp fragment so..
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*Gel extracted the 1500bp fragment from the PCR sample
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*I began sequencing it (Sequencing was finished by David and Kelly). Will take samples to MBSU tomorrow

Revision as of 16:43, 8 August 2008

Saima

  • PCRed PCAM2201 using Cam35s Forward and Reverse Primers. (Protocol to be followed is given in the Masterbook)
    • Digested the PCR product with Xba1/Pst.Ran them on 1%gel. Gel purified the bands.
      • Ligated the gel purified product with J61003.

Chris

  • PCR'd Tryp out of pUC57 (the original that Mike supplied)...buuut I used Vf and Vr to do it. I shouldnt have gotten any results because pUC57 doesnt have sites for those primers to bind BUT I did get a band about the right size...nevertheless, whatever was amplified wouldnt have the biobrick suffix and prefix in it so it was discarded.
  • Redid the PCR with the correct primers....got a good, sharp band ~1500bp in size BUT the same band was in the negative control. I didnt want to risk using something that was just a random 1500bp fragment so..
  • Gel extracted the 1500bp fragment from the PCR sample
  • I began sequencing it (Sequencing was finished by David and Kelly). Will take samples to MBSU tomorrow