Cloning hix fragments
From 2006.igem.org
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Transform JM109 cells, along with the pAra-reverse ligations (take out one tube of 200 ul cells and divide it among the transformations, probably 200/6 = 33 ul each) | Transform JM109 cells, along with the pAra-reverse ligations (take out one tube of 200 ul cells and divide it among the transformations, probably 200/6 = 33 ul each) | ||
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| + | We set up the ligations and let them sit at room temp for 5 minutes. Then we realized that we didn't have the primers to perform the pAra-reverse. We put the ligated plasmids into the freezer to avoid wasting JM109 cells. Tomorrow, Trevor hopes to have the pAra-reverse ready assuming the primers come, then hopefully the transformations will be performed by the end of the day. | ||
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| + | -Adam | ||
Latest revision as of 18:24, 20 June 2006
Set up four ligations as follows, with vector only and vector with each of the three annealed hix oligos from the right freezer:
4 ul of the purified EcorI/PstI digested vector from pTet-GFP 2 ul annealed hixL, hix R, hixC, or H2O 3 ul H2O 10 ul 2X Quick ligase buffer 1 ul Quick T4 ligase
room temp 5 min
Transform JM109 cells, along with the pAra-reverse ligations (take out one tube of 200 ul cells and divide it among the transformations, probably 200/6 = 33 ul each)
We set up the ligations and let them sit at room temp for 5 minutes. Then we realized that we didn't have the primers to perform the pAra-reverse. We put the ligated plasmids into the freezer to avoid wasting JM109 cells. Tomorrow, Trevor hopes to have the pAra-reverse ready assuming the primers come, then hopefully the transformations will be performed by the end of the day.
-Adam
