Exam 1 review - Cloning: talking about plasmid vectors,...

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Cloning: talking about plasmid vectors, which are used to replicate a specific DNA (they replicate inside a bacterium, inserting a DNA of interest). Used to propagate and generate many copies of a specific sequence. o Genomic DNA (human, artificial) o cDNA (complimentary DNA…complimentary to mRNA) cDNA is good, because it will NOT have introns. Would cDNA be useful out of a prokaryote? NO: because it’s redundant to take out the mRNA and convert it to cDNA PCR: in-vitro process. Important to know at least a little bit of sequence; one can’t go through it if one knows nothing about the sequence. Know the area within the “primer” area (in between the arrows: which is about 20 bp) . o One would melt the strand of DNA first (about 94° C). o Then, you anneal (you have base pairing of the primers to the “template DNA.” Basically, this is allowing re-naturation (cool to 55° C). Adding excess of primer will allow the primer to bind to the DNA, instead of the original DNA binding to itself). o
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This note was uploaded on 04/17/2008 for the course PCB 3063 taught by Professor Marta during the Spring '08 term at University of Florida.

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