From 2010.igem.org
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| - | {{Team:Alberta/Head}}
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| - | {{Team:Alberta/beginMainContent}}
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| - | Protocol 17: PCR
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| - | '''Before you start:'''
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| - | * KEEP EVERYTHING ON ICE. Put Taq back into freezer as soon as you`re done with it. DON`T put back dNTP tubes.
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| - | * Reserve a thermocycler and check what size of tubes it takes.
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| - | * Making a master mix conserves expensive reagents, so try to always use one.
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| - | * You may have to do dilutions of your reagents in order to make them usable for PCR (ex: primers, plasmid DNA...)
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| - | * DON'T PUT PRIMERS OR TEMPLATE INTO THE MASTER MIX. ADD POLYMERASE TO THE MASTER MIX LAST (after your other tubes already have template DNA and primers in them)!
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| - | '''Protocol:'''
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| - | *PCR buffer 5ul
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| - | *10uM dNTP's 1ul
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| - | *50uM MgCl2 2ul
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| - | *forward primer 2.5ul
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| - | *reverse primer 2.5ul
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| - | *1ng template DNA 1ul
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| - | *Taq polymerase 0.5ul
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| - | *MilliQ (H2O) 35.5ul (to make total volume 50ul)
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| - | TOTAL 50ul
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| - | [[Team:Alberta/Notebook/protocols| Back]]
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| - | {{Team:Alberta/endMainContent}}
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Latest revision as of 17:13, 26 October 2010
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