Team:WashU/Week8
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+ | We discovered that our problem with the PCRs was due to not optimizing the amount of magnesium chloride needed for the Taq polymerase. Instead, we had been using the Taq buffer with magnesium chloride already added, which was not suited for the reactions we were running. In the gel picture below, the first three wells contain DNA using the buffer with pre-added magnesium chloride, and the last three wells show what happens when we calculate and manually add the proper amount of MgCl2 to optimize Taq polymerase activity. (Also, Z=ZCD, U=GTCs2, and C=CrtZ.) | ||
+ | <div align="center">https://lh4.googleusercontent.com/-qhlXSIMl1d8/UAhpneZ2cOI/AAAAAAAAAWI/9auR5qDv-d0/s800/pcrmgcl.jpg</div><br> | ||
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This morning, we miniprepped a culture of PC42, our ligation of plasmid PSL2131 and construct CS42S. Then, we ran a gel of double and single digests of PC42, cut with X and P and just P, along with an uncut PC42 as a control. (2x refers to ligation cut with both X and P, 1x indicates ligation cut with P, and 0x signifies uncut PC42) | This morning, we miniprepped a culture of PC42, our ligation of plasmid PSL2131 and construct CS42S. Then, we ran a gel of double and single digests of PC42, cut with X and P and just P, along with an uncut PC42 as a control. (2x refers to ligation cut with both X and P, 1x indicates ligation cut with P, and 0x signifies uncut PC42) | ||
<div align="center">https://lh5.googleusercontent.com/--ulNrdD-1xs/UAhk8NfQmeI/AAAAAAAAAV0/BVhDt7rnPAY/s800/pc42digest%25207-18.jpg</div><br> | <div align="center">https://lh5.googleusercontent.com/--ulNrdD-1xs/UAhk8NfQmeI/AAAAAAAAAV0/BVhDt7rnPAY/s800/pc42digest%25207-18.jpg</div><br> |
Revision as of 20:14, 19 July 2012