sequencing long PCR fragment
Posted by: jelena (IP Logged)
Date: July 08, 2009 10:05PM

Hi,

we want to sequencing 1600 bp PCR fragment but we only have 50 cm capillary and pop-7. Can we sequencing with this?

Re: sequencing long PCR fragment
Posted by: Chromous Sequencing (IP Logged)
Date: July 21, 2009 11:35PM

Hi Jelena,

The 1600 bp PCR fragment has to be first sequenced using the forward and reverse PCR primers followed by a 3rd primer designed based on either the forward or reverse data and finally the data has to be aligned.

Re: sequencing long PCR fragment
Posted by: nucleics (IP Logged)
Date: July 22, 2009 07:27AM

Hi Jelena

You shouldn't need a internal primer if you run your traces through our free LongTrace or PeakTrace software. This should increase the read length over 1000 bases and will give you enough good sequence to get the entire 1600 bases with only the two end reads.

Daniel Tillett
Nucleics Support



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