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Team:The University of Alberta/5 August 2008
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'''Chris''' | '''Chris''' | ||
*Winnie redid the ligation of J6 and Tryp. I put transforming this on hold though because... | *Winnie redid the ligation of J6 and Tryp. I put transforming this on hold though because... | ||
| - | **She also did colony PCR on some more of the previous transformants. Got the same result as the two times I did it before; bands ~600bp. It might be GFP - if it is, then it means the digest of J6 I originally did didn't work. I'm doing PCR on the digest to check; if I get a band it means the digest didnt work and that explains the odd results. Im also doing PCR on some undigested GFP to compare and determine if it is GFP or some other fragment. | + | **She also did colony PCR on some more of the previous transformants. Got the same result as the two times I did it before; bands ~600bp. It might be GFP - if it is, then it means the digest of J6 I originally did didn't work. I'm doing PCR on the digest to check; if I get a band it means the digest didnt work and that explains the odd results. Im also doing PCR on some undigested GFP to compare and determine if it is GFP or some other fragment. '''UPDATE''': Got some strange results. 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''' | '''Saima''' | ||
*Did Colony PCR on Col.4, 5 & 6 of I0500+ER01 and I0500+ER02.Set up the O/N's for each of these colonies. Streaked for single colonies | *Did Colony PCR on Col.4, 5 & 6 of I0500+ER01 and I0500+ER02.Set up the O/N's for each of these colonies. Streaked for single colonies | ||
**Ran them on 1%gel. ER02 band size is not the right one. | **Ran them on 1%gel. ER02 band size is not the right one. | ||
Revision as of 16:28, 8 August 2008
Back to the Lab after the Long Weekend!
Chris
- Winnie redid the ligation of J6 and Tryp. I put transforming this on hold though because...
- She also did colony PCR on some more of the previous transformants. Got the same result as the two times I did it before; bands ~600bp. It might be GFP - if it is, then it means the digest of J6 I originally did didn't work. I'm doing PCR on the digest to check; if I get a band it means the digest didnt work and that explains the odd results. Im also doing PCR on some undigested GFP to compare and determine if it is GFP or some other fragment. UPDATE: Got some strange results. 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
- Did Colony PCR on Col.4, 5 & 6 of I0500+ER01 and I0500+ER02.Set up the O/N's for each of these colonies. Streaked for single colonies
- Ran them on 1%gel. ER02 band size is not the right one.
