20.109(F12): Quick Facts: Difference between revisions
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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? | ||
* 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? | |||
Revision as of 13:47, 8 September 2012
DNA Polymerases
Taq Polymerase
- Taq has an error rate of about 1 in 9000. Reference: an article in the journal Biochemistry, quick info can be found in the abstract.
- 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?
- 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?
