BIOL 1000 Lecture Notes - Lecture 4: Phosphodiester Bond, Multiple Cloning Site, Cloning

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Need to amplify gene that want to study, make lots of copies: need large sample size, need situation where gene can be manipulated. Recombinant dna must be created: dna from two or more different sources that are joined together. Promoter: gene not expressed if not expressed. Gene of interest: want multiple copies in multiple plasmids, increase probability that these two well come together, open up cell and nucleus and take out the gene wanted. Denature the dna (break hb bonds) at 95c. Anneal: drop temp to have dna primer attach. Extend: bring temp back to 75c for dna polymerase. These steps are done over and over again. Machine will control the temps for the cycle. Lots of prep, need to make sure that got gene of interest and got right dna primer. Get clear tubes at end of pcr, how do we know what happened.

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