Last week, after learning the nuances of the lab spectrophotometer, I was able to run my previous week's extractions through the spec and get some readings. According to the data collected, I was successful at extracting DNA. This is excellent news, as far the progression of my research is concerned.
I was quite surprised by the differences in the amount of DNA successfully collected and what materials provided those results. The methodology utilized in both of my extractions was identical, only varying in the type and amount of protein precipitation solution I used to clean up the sample. I used an expensive proteinase K dilution ($40/ 5 mg vial, diluted @ 5 mg/250 µL) for protocol number one and an inexpensive (99 cents, diluted at 0.05 g/µL), commercially available meat tenderizer dilution for protocol number two. I had expected the proteinase K solution to realize the best results, yet instead it was the cheaper solution that produced the most DNA. Protocol number one only showed a DNA concentration of 15 µg/mL, while the cheaper protocol number two yielded 1535 µg/mL. This was a substantial difference that only reminded me that, to date in my research, the simpler methods are often the most useful.
I am still searching for a DNA sample of known concentration that we have in-stock, so that I can dilute it and take a spec reading on it. Since the concentration will already be known, I can use it as a control to verify that the measurements I am taking on the spec are in fact correct. More on that to follow as this project progresses.
Until next week, cheers! And enjoy this Ted talk from one of the Nobel Laureates who discovered the structure of DNA, Dr. James Watson.
http://video.ted.com/talk/podcast/2005/None/JamesWatson_2005-480p.mp4
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