BIOL 5060 Lecture Notes - Lecture 12: Polyacrylamide Gel Electrophoresis, Base Pair, Exonuclease
Document Summary
Last time looked at detecting dna via hybridization. Rna and load it more times to get the loading less saturated. Dna: output- gives you cycle number and the threshold signal the machine is looking to see if your product reaches; graph should show an exponential increase/ a doubling with each cycle. Compare experimental and control measure difference between when the two reach threshold. Control reaches threshold at cycle 23: can then put results through a melt curve in which case the machine keeps the sample to see when the signal is lost by dna melting. The experimental because it gives a greater signal (it takes 3 cycles of pcr to make same amount of product on the control as it did in the experimental) 20 vs. 23 cycles 8 fold more experimental than control: key differences: real time vs. reverse transcript, northern- fine for abundant material good visualization, reverse- better for quantization.
