20.109(F12): Quick Facts

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(Taq Polymerase)
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** So would you use Taq for cloning purposes?
** So would you use Taq for cloning purposes?
** What is the probability of finding a mutation in a GFP gene PCR-ed up using Taq polymerase?
** What is the probability of finding a mutation in a GFP gene PCR-ed up using Taq polymerase?
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* If you had a plasmid of size 3 kb, at a concentration typical of high copy plasmids (~150 ng/µL), and you added 1 µL of DNA into a PCR reaction (50 µL), given the typical conditions for a PCR, what are the concentrations of plasmid and primers respectively?
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* Would the concentrations be sufficient to explain why primers preferentially bind to the template compared to re-annealing of the template? If so, why? If not, why?

Revision as of 16:47, 8 September 2012

DNA Polymerases

Taq Polymerase

  • If you had a plasmid of size 3 kb, at a concentration typical of high copy plasmids (~150 ng/µL), and you added 1 µL of DNA into a PCR reaction (50 µL), given the typical conditions for a PCR, what are the concentrations of plasmid and primers respectively?
  • Would the concentrations be sufficient to explain why primers preferentially bind to the template compared to re-annealing of the template? If so, why? If not, why?
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