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Home Q & A Board Biology-Related Homework Help Genetics and Developmental Biology
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ehd123 ehd123
wrote...
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Posts: 778
Rep: 54 2
7 years ago
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
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B.Sc in Biology
M.Sc Neuroscience
PhD. Candidate in Neuroscience

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wrote...
#1
7 years ago
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.
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