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Hello,
I have an issue with some sequences I got from a miniprep and I would like to have your opinion on it.
I sent the same miniprep for sequencing with 6 different primers that cover a 3 Kbp long insert.
Among these primers 3 give a sequence that partially overlap.
2 are reverse primed, and one is forward primed.
The funny thing is that for a particular nucleotide position, in one orientation I get a clear identity, but in the other one I get a clear but different identity.
Forward position identity (in green) is an A, while reverse (in red) is a T.
I think this cannot be a mixture of plasmids as in that case, the picks would both appear on the sequencing trace as overlapping and of similar intensities.
Among the 5 minipreps, this position reproducibly comes up with this “defect”.
Anyone ever experienced that?
Every feedback is welcome.
Thanks.
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