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Team:UC Davis/Notebook/Week 10
From 2011.igem.org
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Ligated and transformed yesterday's digestion. | Ligated and transformed yesterday's digestion. | ||
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| + | --Wednesday 8/17/11-- | ||
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| + | Once again, their are quite a few colonies on our vector control plates. Fortunately, our ligation involved putting promoters (R0010 and R0040) in front of parts containing GFP (E0240+repressor). This means that a successful ligation/transformation will produce a green colony. There were no green colonies on our vector controls which indicated that any green colonies were successful ligations! We did a quick PCR screen and confirmed that this was the case. We now have the nine parts we need to screen and characterize our mutants! | ||
| + | |||
| + | --Thursday 8/18/11-- | ||
| + | Yesterday we cultured our four completed construction so that we could miniprep them today. | ||
Revision as of 17:38, 25 August 2011
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--Monday 8/15/11--
We got our sequences from last week in and checked them to make sure they looked good. Not all of the parts were correct, but we are about a day or two from being done with construction!
Here is a table of which parts are completed and which parts we need to complete in order to finish construction.
| LacI | c1 | TetR | |
| Repressor Screening | *R0010+E0240+I13453+B0034 | R0051+E0240+I13453+B0034 | *R0040+E0240+I13453+B0034 |
| Promoter Screening | E0240+I13453+B0034+C0012 | E0240+I13453+B0034+C0051 | E0240+I13453+B0034+C0040 |
| Wild Type Controls | *R0010+E0240+I13453+B0034+C0012 | R0051+E0240+I13453+B0034+C0051 | *R0040E0240+I13453+B0034+C0040 |
| ' | LacI | c1 | TetR |
| Repressor Screening | R0010+E0240+I13453+B0034 | R0051+E0240+I13453+B0034 | R0040+E0240+I13453+B0034 |
| Promoter Screening | E0240+I13453+B0034+C0012 | E0240+I13453+B0034+C0051 | E0240+I13453+B0034+C0040 |
| Wild Type Controls | R0010+E0240+I13453+B0034+C0012 | R0051+E0240+I13453+B0034+C0051 | R0040E0240+I13453+B0034+C0040 |
In order to finish our construction, we are going to cut out R0010 and R0040 as inserts and put them in front of the parts needed. Tomorrow We will transform and hopefully have the finished construct by Wednesday.
--Tuesday 8/16/11--
Ligated and transformed yesterday's digestion.
--Wednesday 8/17/11--
Once again, their are quite a few colonies on our vector control plates. Fortunately, our ligation involved putting promoters (R0010 and R0040) in front of parts containing GFP (E0240+repressor). This means that a successful ligation/transformation will produce a green colony. There were no green colonies on our vector controls which indicated that any green colonies were successful ligations! We did a quick PCR screen and confirmed that this was the case. We now have the nine parts we need to screen and characterize our mutants!
--Thursday 8/18/11-- Yesterday we cultured our four completed construction so that we could miniprep them today.
