Hi
Why two different bands appear for PCR products of a same gene, when run them in a same gel, same time (master mix were prepared using same reagents and Cdna). I had quadruplicates in PCR run and when those were run on a gel, bands looked weird. Image attached. Can anyone please advice / share your thoughts on this.
Thanks
Prabha
Hi,
is this exactly the same sample for all 4 lanes? If not does your Gene of interest show different isoforms? Which size would you expect for your amplicon and last question which kind of chemistry did you use to amplify the fragment?
Sven
one thing to add: Did your primers span exon-exon junctions or could the longer product also be a gDNA amplification?
Are either of those bands the size you were expecting from either gDNA or cDNA? Which lanes are your positive and negative controls?
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