BIOL 206 Lecture Notes - Lecture 19: Taq Polymerase, Kary Mullis, Ethidium Bromide
16 Apr 2016
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Understand different applications of gel electrophoresis & pcr in. Method used to separate dna and rna molecule by size. Moving negatively charged nucleic acid molecules through agarose matrix with an electric field (electrophoresis) During polymerization, sugar molecules cross link with each other which causes solution to gel into a semi-solid matrix. Shorter molecules move faster and migrate further than longer ones. Most common dye used make dna and rna bands visible is ethidium bromide (etbr) Load dna samples into at top of gel. Method used to make millions of exact copies of a defined segment of dna from a biological sample. Short piece of single stranded dna that flank the target. Attaches nucleotides on the growing strand of dna. Reactions are placed in a machine, thermal cycler. Heat samples to high temperature (94 degree celsius) for ~1 minute to denature the double stranded template dna. Drop temperature drops to around 50 to 60 degrees celsius to allow primers to anneal.
