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Team:UC Davis/Notebook/Week 7
From 2011.igem.org
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== Week 7 == | == Week 7 == | ||
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--Tuesday 7/26/11-- | --Tuesday 7/26/11-- | ||
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Today was also "Tip Day"! We filled enough tip boxes to last us a quite a while. | Today was also "Tip Day"! We filled enough tip boxes to last us a quite a while. | ||
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--Wednesday 7/27/11-- | --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. | 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-- | --Thursday 7/28/11-- | ||
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As usual we're tinkering with our wiki and constantly updating it with new content. | As usual we're tinkering with our wiki and constantly updating it with new content. | ||
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--Friday 7/29/11-- | --Friday 7/29/11-- | ||
We extracted the digestions from yesterday but got poor concentrations so we did a fast digest so that we could come in over the weekend and hopefully have successful ligations by Monday. | We extracted the digestions from yesterday but got poor concentrations so we did a fast digest so that we could come in over the weekend and hopefully have successful ligations by Monday. | ||
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--Sunday 7/31/11-- | --Sunday 7/31/11-- | ||
We ligated R0040+E0240+I13453 in front of both B0034+C0040 and B0034, and ligated R0051+E0240+I13453 in front of both B0034+C0051 and B0034. We then transformed these products, along with their controls, onto carb plates. | We ligated R0040+E0240+I13453 in front of both B0034+C0040 and B0034, and ligated R0051+E0240+I13453 in front of both B0034+C0051 and B0034. We then transformed these products, along with their controls, onto carb plates. | ||
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Revision as of 00:24, 30 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.
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.
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.
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.
We extracted the digestions from yesterday but got poor concentrations so we did a fast digest so that we could come in over the weekend and hopefully have successful ligations by Monday.
We ligated R0040+E0240+I13453 in front of both B0034+C0040 and B0034, and ligated R0051+E0240+I13453 in front of both B0034+C0051 and B0034. We then transformed these products, along with their controls, onto carb plates.
