Team:The University of Alberta/6 August 2008

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'''Chris'''
'''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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*Got some strange results from the PCR I did yesterday. There was a band in the digested sample BUT it wasnt the size of GFP; rather it was ~1500bp, the expected size for Tryp. Even odder is that the same band is in the uncut sample as well, in addition to the expected GFP sized band. I discussed this with Doug for quite some time and we were unable to come to a conclusion about what's going on with Tryp. I figure tomorrow I'll just go back to the drawing board and start over...
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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
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'''Saima'''
'''Saima'''

Revision as of 16:43, 8 August 2008

Chris

  • Got some strange results from the PCR I did yesterday. There was a band in the digested sample BUT it wasnt the size of GFP; rather it was ~1500bp, the expected size for Tryp. Even odder is that the same band is in the uncut sample as well, in addition to the expected GFP sized band. I discussed this with Doug for quite some time and we were unable to come to a conclusion about what's going on with Tryp. I figure tomorrow I'll just go back to the drawing board and start over...

Saima

  • Mini-preped the O/N's of Col. 4,5,& 6 of I0500+ER01 and I0500+ER02. Can be found in Mini-prep Box 3.
  • Sequenced Minipreps of Col.4 & Col.5 of I0500+ER01, and Col.4 of I0500+ER02. To be taken to MBSU on Thrursday. Results to be back by Friday.
  • Set up the O/N's for I0500+ER01 col.6, I0500+ER02 col.4, I0500 in new vector, GFP+B0034 Col.1a & 1b.
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