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Team:UC Davis/Notebook/Week 7
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We decided it was time to revamp our error-prone PCR protocol. Using [http://www.springerlink.com/content/t701h6021352406w/#section=87105&page=1 this] paper as a guide, we created 16 different protocols, with the intention of picking the one that produces the most viable mutants. | We decided it was time to revamp our error-prone PCR protocol. Using [http://www.springerlink.com/content/t701h6021352406w/#section=87105&page=1 this] paper as a guide, we created 16 different protocols, with the intention of picking the one that produces the most viable mutants. | ||
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| + | --Thursday 7/28/11-- | ||
| + | |||
| + | Today we continued construction. We're almost done with our testing construct and have are making headway with producing our mutants. | ||
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| + | We cut: R0051+E0240+I13453 and R0040+E0240+I13453 with Eco and Spe in order to put B0034+C0051 and B0034+C0040 downstream by cutting with Eco and Xba. We also cut E0240+I13453 and B0034+C0012 so that we could put them together for use in the LacI testing construct. | ||
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| + | As usual we're tinkering with our wiki and constantly updating it with new content. | ||
| + | |||
| + | --Friday 7/29/11-- | ||
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| + | We extracted the digestions from yesterday but got poor concentrations so we did a fast digest with the parts cut in a fashion that would allow us to do a triple resistance ligation this afternoon. | ||
Revision as of 21:58, 12 August 2011
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Week 7
--Monday 7/25/11--
We continued with construction today and, once again, extracted the cut Promoters+GFP and repressors from a gel. We ligated the promoters+GFP in from of I13453 and the repressors before B0034. Hopefully this is the last time we will have to ligate and transform these parts.
--Tuesday 7/26/11--
The insert controls for yesterday's transformations looked good, but there were still a good amount of colonies on the vector controls. We decided to do a PCR screen of these parts to make sure that they were the right length.
Today was also "Tip Day"! We filled enough tip boxes to last us a quite a while.
--Wednesday 7/27/11--
We decided it was time to revamp our error-prone PCR protocol. Using [http://www.springerlink.com/content/t701h6021352406w/#section=87105&page=1 this] paper as a guide, we created 16 different protocols, with the intention of picking the one that produces the most viable mutants.
--Thursday 7/28/11--
Today we continued construction. We're almost done with our testing construct and have are making headway with producing our mutants.
We cut: R0051+E0240+I13453 and R0040+E0240+I13453 with Eco and Spe in order to put B0034+C0051 and B0034+C0040 downstream by cutting with Eco and Xba. We also cut E0240+I13453 and B0034+C0012 so that we could put them together for use in the LacI testing construct.
As usual we're tinkering with our wiki and constantly updating it with new content.
--Friday 7/29/11--
We extracted the digestions from yesterday but got poor concentrations so we did a fast digest with the parts cut in a fashion that would allow us to do a triple resistance ligation this afternoon.
