This is my first post on this forum and I'm looking forward to discussing all sorts of my, and other problems, findings etc!
I've done an RT-PCR on the PML-RARA translocation in the human NB4 APL cell line. I blasted my primers and the expected fragment length should have been 128 bp. I loaded my samples on an 1% agarose gel, along with Molecular weight marker 8 (from Roche I believe. The molecular weight marker is distorted, I accidently put the tip of the pippette into the gel itself when loading the samples.
The problem is, my amplification product, isn't quite 128 bp, but based on the mm8, it is more or less around the 140 bp (quite the difference). I can't really explain why this is, apart from my poor pippeting skills.
I enclosed the pic of the electrophoresis for your viewing pleasure!
Thanks in advance for answers and solutions, and for your interest in this topic!
PS. The RNA was isolated using Tripure reagent and cDNA was made using the cDNA first strand synthesis kit by Roche!