Dear all,
I'm wondering about a possible substitution of a G>A base or a deletion of the G base in view of the chromatograms I've analyzed. I would like to point out that I have observed the same thing in 8 different samples.
Your opinion would be a great help.
Sincerely
Ref seq : GCCTCAGCGCC
Muted seq: GCCTCACGCC
The spacing between the 2 adjacent bases suggests that there is a base (maybe G) present even though the peakintensity is so poor that it could be background. If there was a deletion then all heterozygous samples would be a real mess after the deleted base and this is not the case here. There are a few vry weak intensity G bases ( eg base 82) in this sequence so I think that sequencing the reverse strand should produce a better electropherogram and clear this up, Both base spacing and dye problems happen this early in the sequence so reverse sequencing helps. Can you attach an original sequence .abi file from the company which may have additional information to help troubleshooting please?
The chromatogram quality is not so good, but I can detect a small peak (in black) corresponding to G at this location.
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