Today I will do colony PCR using 1, 5, 8 and 10 DNAs. This is to ensure DNA is going out signals. The gel was wrongly ran at 20V instead of 100V, but that's okay because the whole purpose of this PCR was to ensure that the DNA present is strong enough to give out a signal, and it is, as we can see in the image below.
DAY 14: 27/06/17
Today the only thing I had to do was to plate out the b3.4 1/100 dilution and 3 1/100 dilution grown solutions.
Plate out the solutions were fairly quick, so to do more things in the lab, I helped Dr. MacNeill with his own research buy doing colony PCR with some of his plasmids DNAs.
DAY 15: 28/06/17
Today PCR was ran using 1, 5, 8 and 10 plasmid DNAs with NRP1-NRP2 and NRP3-NRP4 mix primers. Results are shown in figure 7.
As we can see from figure 7, PCR results suggest that the PCR didn't work, so everything was repeated with optimising conditions, until it worked and the DNA gave out signals.
➢ A 'wildtype' was induced and a new primer replaced NRP2 was used: K185-REV primer (figure 8)
➢ Optimising conditions also involved changing the annealing temperature in the PCR reaction (annealing temperature went from 60oC to 48oC) (figure 9)
Results are shown in figure 8 and 9.
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Ok, so Dr MacNeill is going on holidays with his wife, which means, I get some free time myself!!! 😚😃
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