I have sequenced a sample with a custom AmpliSeq pannel. On the ion torrent results I see an already described pathogenic mutation in heterozigosity with 578 reads (285/293). When I do the sanger sequencing to confirm this result I have an heterozigosity, but like 20%/80%.
Can you explain the difference? Do you have any solution? How can I be sure that both alleles are being ampplified in the same amount? I have already tried several different primers.
Thank you all.