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Thursday, November 9, 2017

Results... At Last!

On October 31, 2017, I ran my post-PCR DNA samples from October 26, 2017 under electrophoresis. Theses particular samples were different because I doubled everything, except the DNA. I added 20 microliters of Master Mix, 11 microliters of sterile water, 4 microliters of the forward primer, 4 microliters of the reverse primer and 1 microliter of DNA. All this totaled to an amount of 40 microliters. The primers used were 27F and 1492R. Once I ran these under electrophoresis, I finally got results (Figure 1)! I was so excited because at one point, I was beginning to think I was doing my procedures wrong. Nonetheless, I am happy to say that this change made a difference. Also, this week, I started growing bacteria in TSB broth (Figure 2) that was isolated by Samantha Faltermeier, an S-STEM intern. I will be performing DNA extraction and, eventually, PCR on these unknown bacteria!
Figure 1. The resulting gel after it was ran under electrophoresis. The post-PCR DNA in each row from left to right is as follows: MW Ruler, 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 & E. Coli. The particular DNA that amplified were 4, 9, 10, 11, 12 & 13. These were the ones that showed bright bands.

Figure 2. Unknown DNA samples from isolated bacteria in a Creosote bush. The samples were transferred from a TSA plate to these TSB broths. They are growing at room temperature and the samples are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 & 14.

1 comment:

  1. Great job on getting results for your Post-PCR DNA samples! I definitely struggle with constantly wondering if I am running tests right or wrong. This past week, when all nine of my samples were gram positive, I thought I had to have done something wrong, especially since I struggle with getting good results for my gram stains. Anyways, good luck on your next step with the Creosote bacteria samples!

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