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Genewiz Sequencing Problem

no priming

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Posted 14 June 2012 - 06:25 AM

Hi, I have submitted a plasmid to Genewiz for sequencing on 3 occasions. All three times the plasmid was not sequenced due to "no priming". The only thing they tell me is that it can be a concentration problem or contamination. The plasmid I have sent has come from miniprep's and maxiprep's and looks identical to the original plasmid when run on a gel. I have used universal primers from Genewiz (T7 and Neomycin) and neither worked due to no priming (the plasmid has these sites). Has anyone had a similar problem and how did you overcome it? Any advice would be greatly appreciated.

Side note. I have linearized the plasmid in separate reactions just fine. So I doubt there are inhibitors in the preps.


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