Introns and RT-PCR - RT-PCR and Escherichia coli (Oct/15/2003 )
I wish to perform RT-PCR using primers specific to E.coli genes. However as E. coli lacks introns, can I assume I can use my normal primers that I use for PCR, as long as I have removed all traces of DNA from my isolated RNA. Hope someone can clarify this for me.
when you say RT-PCR, do you mean real-time PCR of reverse transcrition PCR? with real time you can simply treat your RNA with DNAse prior to reverse transcription and that will fix your problem of not having primers that distinguish between genomic and cDNA. you do experience some loss in the subsequent purification steps, but it's the best way to go.