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Title: RT PCR ampliified cdna Post by: ehd123 on Dec 18, 2016 performing a reverse transcription PCR experiment.. how can i determine whether my bands correspond specifically to amplified cDNA?
a) design primers that flank introns b) amplify unreverse transcribed RNA c) add a probe that hybridizes to my amplicon d) rnase my sample e) a and b f) a and d g) b and c Title: Re: RT PCR ampliified cdna Post by: akorne on Mar 4, 2017 A: design primers that flank introns.
If you have genomic contamination that you amplify then the intron will be included and your product will be too large. B is wrong because RNA must be reverse transcribed before PCR C is possible but an extraordinary waste of time as you will need to do a Southern blot which means you will run a gel and be able to see the size of the product at this stage. D is wrong as this is an integral part of the RT-PCR method. |