Attempted PCR using the P11/P12 primers and thermocycler parameters described in the previous entry. This is the first time I have used these primers and am using a program that I designed myself. Amplicons were mixed with EZ vision loading dye using two different volumes of DNA since I had been having issues with overloading the wells in the past. Either 5 uL or 10 uL of amplicon DNA was combined with 2 uL of the loading dye and added to the wells of a 1% gel. The gel was ran for 40 minutes at 100 volts. Below is the resulting gel with labels below that:
Lane 1 | Lane 2 | Lane 3 | Lane 4 | Lane 5 | Lane 6 | Lane 7 | Lane 8 |
Hi Lo | Transcon | pMarA | 9545 | Transcon | pMarA | 9545 | Empty |
Lanes 2 - 4 were the 5 uL volumes of DNA and Lanes 5 - 7 were the 10 uL volumes.
Discussion:
I may repeat this amplification test using the other transconjugate I have acquired in the future, however it is more likely I will re-attempt the tripartite conjugation and makes changes with the time intervals in hopes of improving my results.
//EWW
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