Company supplied just forward primer for Sanger sequencing. I guess, there is no needs to amplification inserts using PCR thermocycler because it was done by bacteria itself. Now, should I begin sample preparation from BigDye Terminator step with ignoring amplification target fragments (inserts) and first purification?
Hi Mahdi Shekari ,
If I understand your question correctly, you have been supplied with a purified plasmid with your insert already cloned in? If so, just confirm the appropriate concentration needed and proceed with BigDye with the primer supplied.
It will depend upon your insert size, if your insert size is small, then you can directly sequence with the primers provided but if you have a larger insert you might want to design more than one set of primers.
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