MadSci Network: Molecular Biology |
If you are planning to use in vitro transcription to produce the mRNA from your gene of interest, you will need a clone which contains the promoter and the complete ORF, and preferably little else. So you would need the complete sequence to generate the ideal clone. There are a couple of ways I can imagine to do this. 1. Use the sequence of your partial clone to probe a cosmid/plasmid library and then sequence the insert of positive clones. 2. By an arbitrary primed PCR approach. This could be messy. The short answer, if I understand your question correctly, is that I can't think of a way to specifically produce mRNA from your gene without knowing the DNA sequence. If I misunderstood, please rephrase the question and post again.
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