BIL 250 Lecture Notes - Lecture 9: Kary Mullis, Polyacrylamide Gel Electrophoresis, Real-Time Polymerase Chain Reaction
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Pcr was just cheaper yo by 10 fold. Enabled amplification of millions of copies of dna. Performed in thermal cycler, but first used heat blocks of different temperatures and moved the tubes around. Pcr can run as high as 70 c. You can cycle through as many aas 40 heat cycles. Takes two primers and they polymerize the dna. We do a last minute extension to extend any incomplete products. Double strand temp raises to denature bond between two strands. Temp is raised so the tac polymerase extends. You generally can"t visualize the dna until after 20 seconds. Don"t fall of the template and have a low error rate. Incorporate a dye that flouresces when it"s bound to dna. Useful in ecology of a heterogenous sample to measure the relative abundance. You have to measure the amount as it is produced. The more green, the more is produced. Do it in a optical tube or plate.