Hello, Any suggestions on how to obtain the sequence of beta-lactamase a & w genes in the plasmids. This plasmids have been donated to our lab and now we want to sequence them to develop an assay and we really have no idea about the back bone. We initially tried to sequence them with pCMV sequencing primers, we didn't get any sequence. Can anyone provide me suggestions on what would be a best way to obtain the sequence on the plasmids.
Look at the sequence of traditional vectors like pUC, pGEM, pET, pCDNA because the beta-lactamase genes were in most cases identical. If T3 and T7, or M13 primers are not working try a primer sequence from the above mentioned sources.
To test with T3, T7 or M13 primers as Michael suggested is a good idea. Note, however, the different variations in sequence of the different M13 primers typically used. For an overview, see http://openwetware.org/wiki/Common_primer_sequences.
Thanks you very much for the inputs, Ok I will try the sequence using the suggested sequencing primers.
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