PCR variability or bad cell source? - (Oct/18/2007 )
here is what I am struggling with PCR and I would really appreciate if I can get any feedback on this.
I have human cells and passaged them for a total of 7 samples. I had quite some trouble in hacing the right cDNA, but now I seem to have it as my gene control (b-actin) is showing up fine.
My problem is that when I run other markers specific for the cell population I almost get nothing. Out of 4 markers I tried so far, I have had very faint bands for one marker only and for 2 out of 7 samples for a second marker.
I know it might be an issue with primers, but I have used these primers with other cDNA few weeks ago and they work just fine.
Try with fresh aliquot of solutions for the PCR. Like get new dNTP, to preparing new primers solutions. These could help.
Good Luck !!!