Week 13- Elusive results

Last week, I was able to run another PCR and electrophoresis gel on my E. coli DNA samples. This time, I changed the primers that I paired to two that had melting temperatures with 4°C of each other, due to the prior PCR's annealing temperature being too low to generate results.

These are the primers that I used: 27F (AGAGTTTGATCMTGGCTCAG) and 1492R (GGTTACCTTGTTACGACTT). They had melting temperatures of 59.4°C and 55.8°C, respectively, so I ran the PCR with an annealing temperature of 54°C.

Unfortunately, while there was DNA present in my gel, it was sheared and did not successfully amplify. Here's a screen capture of the first gel from last week:

And here is the secondary image with the higher annealing temperature of 54°C:

As you can see, both gels are lacking the clearly define horizontal bands that indicate the presence of amplified DNA. So I will have to refine and repeat my methodology as I proceed. There is good news, however- I have been approved to return as an unpaid intern over the summer session. I will be able to continue my research. I am doing this for the sheer joy of the learning involved, and also so that I will hopefully be ahead in my project with the onset of the fall semester.