Sunday, July 8, 2007
Sequences available!
On Monday, I got the plates I sent off to purification back from the robot on 3rd floor of the Biology building that purifies and sequences plates for the labs. When I used the spec machine to check out my DNA concentration after the purification, nearly all the tubes had a negative concentration! (not good if you want to sequence anything) Unless, of course, the DNA is simply too diluted for the spectrophotometer to read. In that case, there is hope and all is not lost. So I ran a gel on 5 ul of some PCR product comparing it to 1 ul of the stuff I had previously purified myself without using the robot. That way, even if the PCR product was really dilute, I should still be able to see a band. The wierd thing is that all of Lisa's other purifications that were purified along with my samples had no problems with the DNA concentration. At first I thought it might be our spec machine, but I checked some things I had previously purified and they were fine. It's a mystery... So the gel came back with lots of single bands-- meaning my PCR product was in there all along and could be used for sequencing. So I then prepared the plates for sequencing using Big Dye and buffer, but no water since the DNA was already so dilute. That part was easy. When the sequences came back, I got to analyze. What I did on Friday was rename all of the sequences so that their names fit which primer and individual it represents, rather than just a random number. Then I organized the sequences into primer groups. I also got a sneak peek at some SNPs, which were obvious by their characteristic double hump.... more on that next week.
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