IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/25: Difference between revisions
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Last week we had made cultures of V9, V10, V11 and V12 vectors and stored pellets of the resulting cells over the weekend. | Last week we had made cultures of V9, V10, V11 and V12 vectors and stored pellets of the resulting cells over the weekend. | ||
We miniprepped the cells to obtain the plasmids. We did so using the Qiaquick Miniprep kit. As an initial test that we had obtained the plasmid we expected we digested using EcoI and HindIII and ran on a gel. The gel (shown below) | We miniprepped the cells to obtain the plasmids. We did so using the Qiaquick Miniprep kit. As an initial test that we had obtained the plasmid we expected we digested using EcoI and HindIII and ran on a gel. The gel (shown below) showed the presence of the expected bands. | ||
To verify that we had inserted the sequence that we expected into the plasmid we planned on sending them for sequencing by GeneWiz. GeneWiz require concentrations of 80ng/uL. Our nanodrop results showed that we had obtained low concentrations from our miniprep (<44ng/uL). To achieve higher concentrations of plasmid we re-picked colonies and incubated cultures for longer. | To verify that we had inserted the sequence that we expected into the plasmid we planned on sending them for sequencing by GeneWiz. GeneWiz require concentrations of 80ng/uL. Our nanodrop results showed that we had obtained low concentrations from our miniprep (<44ng/uL). To achieve higher concentrations of plasmid we re-picked colonies and incubated cultures for longer. We miniprepped the cultures and obtained higher concentrations of the plasmid. | ||
We did not have a large enough volume of plasmid for sequencing so we transformed the miniprepped plasmid back into E. coli and grew on LB + Kanamycin. We picked colonies and grew cultures. We obtained the plasmid using a miniprep. | |||
==Team Flavor== | ==Team Flavor== | ||
Revision as of 10:51, 25 June 2010
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Week 2 Summary (6/25/2010)Team VectorLast week we had made cultures of V9, V10, V11 and V12 vectors and stored pellets of the resulting cells over the weekend. We miniprepped the cells to obtain the plasmids. We did so using the Qiaquick Miniprep kit. As an initial test that we had obtained the plasmid we expected we digested using EcoI and HindIII and ran on a gel. The gel (shown below) showed the presence of the expected bands. To verify that we had inserted the sequence that we expected into the plasmid we planned on sending them for sequencing by GeneWiz. GeneWiz require concentrations of 80ng/uL. Our nanodrop results showed that we had obtained low concentrations from our miniprep (<44ng/uL). To achieve higher concentrations of plasmid we re-picked colonies and incubated cultures for longer. We miniprepped the cultures and obtained higher concentrations of the plasmid. We did not have a large enough volume of plasmid for sequencing so we transformed the miniprepped plasmid back into E. coli and grew on LB + Kanamycin. We picked colonies and grew cultures. We obtained the plasmid using a miniprep. Team FlavorTeam AllergyTeam Fence |
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