Day 2 Frustration

Well, today was sort of an off day. I'm still shaking off the rust from last year's winterim and it showed. Today, I set up my first PCR experiment of this winterim. Even while I was doing it I felt like I was forgetting something. However, I continued and eventually tried to analyze my data.

Boy did I make a mistake. This whole process works because we can put a known number of competitor molecules in the PCR and then see the ratio of native (the strands we want) to the competitor molecules. However, based upon my data, I forgot to put the competitor in.

 

(how I felt)

But I realized that it was still just the second day, and that I was still performing tenfold better than how I was at this time last winterim. Plus, I was happy that I was able to troubleshoot and see my own errors. Tomorrow I hope to move past my blunder and start getting some data.

BUT! WAIT! THERE'S MORE!

Xiolu was showing me a different type of PCR: Real Time PCR.

Basically, what this method of getting data gets the same conclusion in a different fashion. It involves measuring the amount of fluorescence during each cycle of PCR. Although PCR occurs as a exponential function, the process eventually exhausts its products, so it reaches a limit. What they do it take the threshold at which the Native and the Competitor Templates reach their limits and then use this information to formulate a exponential expression that can be used to determine the ratio of CT to NT. And as I mentioned earlier, since we know the amount of Competitor, we can approximate the number of Natives made during PCR. This is basically the same result as what I've done and am currently doing, but it just can be used for different circumstances.

STILL NOT DONE. I also talked to Doctor Willey about the significance of the project I'm helping Xiolu. What he told is that since they are looking at how does CEBPG affect the transcription of the other genes important in keeping epithelial cells in the airways of the lung non-cancerous. Ultimately, they want to see how important it is in general. So hopefully I can help :)