I am hypothetically looking at a genetic disease arising from a trinucleotide repeat expansion (30 base pairs in total) after the 150’th nucleotide of a 450 nucleotide gene.
The question asks you to design a PCR-based method to detect this disorder We learned about qPCR and gel electrophoresis.
GE: I was thinking that after PCR you could use gel electrophoresis to distinguish the size difference between the amplified product using the same primers. But at the same time I don’t know if a 30 base pair difference is enough to be shown on a gel
qPCR: But then we also talked about how qPCR utilizes the fact that each cycle doubles the of copies of the target DNA and allows you to quantify the amount of transcript in treated vs untreated cells (done by looking at SYBR green fluorescent emission at the end of each extension stage
Which method makes sense?