I'm working on antibiotic resistance and I need to sequence for beta lactamase resistance coding gene. Microbes, in particular. It should be carried out in rt pcr or multiplex pcr and the primer has to be designed for it. Can any one suggest where it can be done. I'm in banglore.
If I understand correctly, then I would suppose RT-PCR may not be needed. Search on NCBI Gene at http://www.ncbi.nlm.nih.gov/gene for the microbe-specific beta lactamase resistance gene. Use the sequence to design specific primers using the Primer3Plus software at http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi/ or the primer design tool on NCBI at http://www.ncbi.nlm.nih.gov/tools/primer-blast/.
Then run DNA extraction, PCR, gel. You may purify the amplicon, and sequence. I suppose multiplexing can be relevant if you are dealing with multiple resistance genes and you need to amplify them together. In that case, the primers have to be in harmony with each other, without the possibility of primer-primer dimers.
I hope this helps.
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