end-point PCR

[ifhmn]

how can I get a consistent result of my PCR reaction. I'm using the same cDNA, primers, and other reagents, but when I repeated again my PCR and I get an extra bands (unknown) on gel electrophoresis and sometimes the bands are not clear as the 1st time I ran my PCR.

[relaxin]

After you thawed out you sample and reagents, did you vortex them to ensure they are homogenous before dispensing?

[ifhmn]

I just thawed them for more than hour without centifuged them. Is this might be the reason that I got different results?

[relaxin]

When a solution is frozen and then thawed, you will see water is on the top while salts and proteins tend to stay in the bottom. You need to vortex the tube to make the solution homogeneous again. Centrifugation is only needed when some solution got into the inside of the cap.