Hi everyone,
I'm doing amplification and purification of 4 different genomic regions. 3 of them are ok, but I'm struggling with my last amplicon. The amplicon length on the gel changes before and after the purification. Before purifying I have a 360 bp and after the purification I have a strong lower band of 250 and a upper band around 400 pb. It cannot be caused by a higher DNA yeld before the purification because I loaded different and small volumes on the gel to be sure about the size. It happened the same with a different primer set for the same region. Thinking about an issue with my primer set I changed it, but the problem reappeared.
Is it possible that somehow the beads interfere with the size of the amplicon? This problem never happened with other genes so I really don't understand what's going on.
Thank you for any suggestion!
Elena
I'm doing amplification and purification of 4 different genomic regions. 3 of them are ok, but I'm struggling with my last amplicon. The amplicon length on the gel changes before and after the purification. Before purifying I have a 360 bp and after the purification I have a strong lower band of 250 and a upper band around 400 pb. It cannot be caused by a higher DNA yeld before the purification because I loaded different and small volumes on the gel to be sure about the size. It happened the same with a different primer set for the same region. Thinking about an issue with my primer set I changed it, but the problem reappeared.
Is it possible that somehow the beads interfere with the size of the amplicon? This problem never happened with other genes so I really don't understand what's going on.
Thank you for any suggestion!
Elena
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