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Team:Technion/4 September 2012
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- Restriction digest of the pCP backbone, Fus2 and Fus2del inserts with XhoI and XbaI.<br> | - Restriction digest of the pCP backbone, Fus2 and Fus2del inserts with XhoI and XbaI.<br> | ||
- Ran the pCP restriction products on a gel. The bands were very weak, which was unexpected because I took 3 ug of DNA. The nanodrop must have had an error when I measured the concentrations before the restriction.<br> | - Ran the pCP restriction products on a gel. The bands were very weak, which was unexpected because I took 3 ug of DNA. The nanodrop must have had an error when I measured the concentrations before the restriction.<br> | ||
| - | - Gave all the miniprep products I have of pCP to Aya, she will concentrate it using the speed vac. | + | - Gave all the miniprep products I have of pCP to Aya, she will concentrate it using the speed vac.<br> |
| + | - Planned with Hila and Roee the set of reaction conditions for the assembly of two phage fragments. We will use 100, 250 and 400 bp overlaps with different incubation times with a T5 exonuclease in 37 degrees prior to the incubation with Gibson mix from NEB. In parallel, we will try using just the Gibson mix in the same reaction conditions (incubation in 37 degrees before the Gibson protocol). | ||
==Inbal== | ==Inbal== | ||
Revision as of 05:16, 5 September 2012
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Ilya
- Ran 5ul of the PCR products on a gel, got the expected bands.
- Purified the PCR products and measured concentration.
- Restriction digest of the pCP backbone, Fus2 and Fus2del inserts with XhoI and XbaI.
- Ran the pCP restriction products on a gel. The bands were very weak, which was unexpected because I took 3 ug of DNA. The nanodrop must have had an error when I measured the concentrations before the restriction.
- Gave all the miniprep products I have of pCP to Aya, she will concentrate it using the speed vac.
- Planned with Hila and Roee the set of reaction conditions for the assembly of two phage fragments. We will use 100, 250 and 400 bp overlaps with different incubation times with a T5 exonuclease in 37 degrees prior to the incubation with Gibson mix from NEB. In parallel, we will try using just the Gibson mix in the same reaction conditions (incubation in 37 degrees before the Gibson protocol).
Inbal
Asaf
I made a fusion PCR between the RiboSwitch and the different polymerase genes with a Tm of 69C. It didn't work as you can see in the gel. Maybe the problem is with the Tm, so tomorrow I will try to find the right Tm using gradient PCR.
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