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Team:Technion/10 September 2012
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I ran the 56C Riboswitch+different polymerase on an extraction gel. I ran T7 of both 55 and 56C because I wasn't sure if he had a 3.2 kb band.<br> | I ran the 56C Riboswitch+different polymerase on an extraction gel. I ran T7 of both 55 and 56C because I wasn't sure if he had a 3.2 kb band.<br> | ||
<gallery perrow="4" widths=150px heights=150px style="color: black; border: 5px ridge #A7A3BF; font: 13px david; background: azure"> | <gallery perrow="4" widths=150px heights=150px style="color: black; border: 5px ridge #A7A3BF; font: 13px david; background: azure"> | ||
| - | Image:10.9_fusion_RS+ | + | Image:10.9_fusion_RS+poly_grad_extraction_gel-view.jpg |
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Revision as of 11:38, 15 September 2012
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Ilya
- Did colony PCR for the pCP+RS+mCherry and pCP+RBS+mCherry clones that Rachel prepared for me. There were some positive results but then I realized that they are useless. The previous insert in the plasmid was also mCherry, so I had no way of telling the difference between the background clones and my clones judging by colony PCR.
- Put starters of the colonies I picked for the colony PCR in order to do a test with BamHI digestion tomorrow to find the correct clones.
- Planned some aspects of the gibson attempts that we are going to do this week with Roee and Hila. Tomorrow we will try the assembly of fragments 4 and 5 with 100bp and 400 bp overlaps with different incubation times in 37 degrees followed by the incubation in 50 degrees which is instructed in the gibson protocol.
- Prepared a 0.6% gel for testing the gel electrophoresis conditions for the gibson results.
Inbal
Asaf
I ran the fusion PCR of the Riboswitch with the different polymerases on a gel. This time I got the required bands (3.2 kb)
and also 2.7kb and 500 bp bands.
 |
I ran the 56C Riboswitch+different polymerase on an extraction gel. I ran T7 of both 55 and 56C because I wasn't sure if he had a 3.2 kb band.
 |
Hila
Lior
Noa
Evgeni
- Due to continuous failure of Bba_015 insertion in pPROLar plasmid I'll try new strategy: instead of double digestion of pPROLar I'll do sequential digestion: HindIII-HF restriction -> Heat inactivation of HindIII-HF at 80C -> BamHI-HF restriction.
- In addition I'll do new Bba_015 restriction with BamHI+HindIII, maybe ligation problems were due to problem with biobrick restriction (Done by Rachel)
- Overnight ligation of newly prepared pPROLar and Bba_015
