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Imperial College/25 September 2008
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[[Image:Minidig25-91.png|center|700px|thumb| U = undigested, D = digested]] | [[Image:Minidig25-91.png|center|700px|thumb| U = undigested, D = digested]] | ||
| - | *All the inserts for | + | *All the inserts for EpsE (above) and 5' AmyE (below) appear correct. One of the inserts for 3' AmyE is a different size to the others. Tomorrow 5' and 3' AmyE minipreps will be cut ''Xba''I - ''Spe''I to check for correct orientation. That gel will be run longer with RFP-Terminator (the possible contaminant) to determine if that may be the cause of the size change |
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*EpsE Single Colony PCR reactions were cut with ''Hind''III to check that the insert was EpsE and not the possible contaminant, RFP-Terminator | *EpsE Single Colony PCR reactions were cut with ''Hind''III to check that the insert was EpsE and not the possible contaminant, RFP-Terminator | ||
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[[Image:Minidig25-92.png|center|700px|thumb| U = undigested, D = digested. EpsE on the left is cut with ''Hind''III, 5' AmyE cut with ''Eco''RI and ''Pst''I]] | [[Image:Minidig25-92.png|center|700px|thumb| U = undigested, D = digested. EpsE on the left is cut with ''Hind''III, 5' AmyE cut with ''Eco''RI and ''Pst''I]] | ||
*The insert in minipreps one and two (numbered left to right) is has lost a small fragment (~150bp) when cut, as should be the case with EpsE which possess a ''Hind''III site about 100-150 away from the 3' end of the gene. The third miniprep shows what appears to be two fragments (one approximately 250bp, the other approximately 400bp), though it is more likely that there are three, as would be the case if RFP-Terminator was the insert instead. | *The insert in minipreps one and two (numbered left to right) is has lost a small fragment (~150bp) when cut, as should be the case with EpsE which possess a ''Hind''III site about 100-150 away from the 3' end of the gene. The third miniprep shows what appears to be two fragments (one approximately 250bp, the other approximately 400bp), though it is more likely that there are three, as would be the case if RFP-Terminator was the insert instead. | ||
Revision as of 00:48, 29 September 2008
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25 September 2008Wet LabCloning
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