Hello,
I use a commercially available real-time PCR kit for poultry virus detection.
I have used this kit for some preliminary experiments but need the primer sequences for further research.
Is it possible to reverse engineer this kit to get the above. Can the primer master mix be purified to extract the oligos so that they could be sequenced via Sanger.
Is the above possible? Any suggestions for processing and method development.
Thanks,
If knowing the 5' end of each primer would suffice, you could clone your PCR fragment into a vector (eg. TOPO-vector), then sequence this insert. However, it would not allow you to know the length of the primer and its full sequence (as you wouldn't know where the primer-part of the sequence ends).
Hi Alexander, why can't the primers be sequenced directly? Is it because of their small length?
Further to Alexanders suggestion, I would just submit the sequence obtained to any standard primer/probe generating program. If the kit was based on an optimised primer/probe set this will most likely have been generated automatically in the first instance so resubmitting the sequence should yield similar results.
Alternatively, you could just design your own primer/probe sets.
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