What does PCR stand for, again?

June 29, 2017

There is a reason that scientists are known for researching, and not just searching. Alas, as with any good research project, we have been struck with the woes of research recently, in that we have hit a roadblock that discontinues the progress we can make on the project. A very successful mRNA extraction was carried out, and a reverse transcription polymerase chain reaction (RT-PCR) was set up, but with no amplification.

 

Enter: troubleshooting. We backed up one step, maybe the primers were not annealing correctly; we performed a gradient RT-PCR to see if an annealing temperature could be elucidated. No luck. Back up once again, check for intact template, and this appeared to be where the issue lay. To test the viability of the primers we are using, we performed a PCR on DNA rather than RNA (since it is moderately easier), but the DNA did not appear to even be there. We attempted to re-extract, but a protocol that has worked countless times before has suddenly stopped working. This is one of those times where I am very grateful for a mentor who is ready to jump into the lab and figure out the problem alongside of myself and another student. So side by side, we will perform the extraction, and I will hopefully learn the technique that will make this whole project proceed once again.

 

Today Dr. Sheridan was addressing the lab and reminded us all of something very important in the field of research. What he said distilled into something along the lines of “if we knew how to do it, we would have done it and already have the answers.” He very eloquently reminded us that research is hard, and the fact that our projects sometimes come to standstills is perfectly fine. It might even be expected. It was perfectly timed, and reminded me to be excited about the results that are hopefully just around the corner, just waiting to be discovered.

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