Wednesday, May 31, 2006

PCR

I finally got the desired product. Yields from 2 mM and 1.75 mM reactions were about equal. I used 50ng of genomic DNA in each and 0.5 uM of each primer, with a final volume of 50 ul. I tried one tube with 100ng of genomic DNA but didn't have a very good product. Program was hotstart 25 cycles of [1min at 94, 2min at 50, 3min at 72] with an initial denaturation and final extension. The bands were fairly faint, so I'm going to try increasing the program length to 40 cycles.

I'm going to take all my posts from the period when I was using a bad primer offline. If anyone desires to see them, contact me.

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