I purified my three PCR products, their concentrations were between 40-60 ng/ųL. Is there any reason why the sanger sequencing of my samples if failed? In the Gel From lift to right first two samples and the 3rd one is the control pcr product. Needs suggestions
Thank you
What method did you use to clean the pcr product from the primers or gel. How much template dna and primer did you use in the sequencing reaction?
Can you attach an actual sequence file from the sequencing company ( .abi file) as it makes troubleshooting much more accurate. Have you recently had any good results of sequencing with the sequencing kit used or do you send template and primer to a commercial company for sequencing?
Dear@Paul Rutland The method I used is membrane base purification kit from "Solars Bio" that require PCR product from the Primers. The Template DNA is 4 ųL and a Primer of 2 ųl each (forward and reverse) of the 10 P/mol. Well i got my sequencing results from Macrogen south korea. (.abi files ) were present but i just extracted pdf files from ZIP document. Yes i got good results of my sequencing from my ist batch samples purified through this kit.
Regarding primer and template, i just provide them my Primer set.
can you attach one of the poor quality .abi files please Salman Hassan
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