Corum:PCR: Difference between revisions
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'''Relevant papers and books''' | '''Relevant papers and books''' | ||
<!-- If this protocol has papers or books associated with it, list those references here.--> | <!-- If this protocol has papers or books associated with it, list those references here.--> | ||
</biblio>--> | </biblio>--> | ||
<!-- Try the [[Template:FormatRef|FormatRef template]]--> | <!-- Try the [[Template:FormatRef|FormatRef template]]--> | ||
#{{FormatRef| | #{{FormatRef|Sambrook, J and Russell, DW|2001|Molecular Cloning: A Laboratory Manual Volume II|Cold Spring Harbor Press, Cold Spring Harbor, NY|8.18-8.24|}} ISBN 0879697164gh | ||
#{{FormatRef|Jacob, F and Monod, J J|1961| |Mol Biol 3(3)|318-56| }} PMID 13718526 | #{{FormatRef|Jacob, F and Monod, J J|1961| |Mol Biol 3(3)|318-56| }} PMID 13718526 | ||
#{{FormatRef|Ptashne, M|2004|Genetic Switch: Phage Lambda Revisited| | |Cold Spring Harbor Laboratory Press}} ISBN | #{{FormatRef|Ptashne, M|2004|Genetic Switch: Phage Lambda Revisited| | |Cold Spring Harbor Laboratory Press}} ISBN 0879695773 | ||
==Contact== | ==Contact== | ||
Revision as of 14:38, 18 June 2012
Overview
Standard polymerase chain reaction (PCR) protocol.
Materials
For a 50 μL PCR reaction:
- 35 μL H2O
- 5 μL 10X PCR buffer
- 5 μL 2mM dNTPs (each)
- 1.5 μL 50mM MgCl2
- 1 μL 50μM sense primer
- 1 μL 50μM antisense primer
- 1 μL 5nM DNA template
- 0.5 μL TAQ DNA polyermerase
Procedure
- In a PCR tube, mix the components on ice in the order they are listed above.
- Perform thermocycling program
- 95 °C 5 min
- 95 °C 30 s
- TH 30 s
- 72 °C 1 min for each 1 kb PCR product
- Repeat steps 2-4 a total of 12-36 times (24 is standard).
- 72 °C 5 min
- 12 °C hold
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
References
Relevant papers and books </biblio>-->
- Sambrook, J and Russell, DW (2001) Molecular Cloning: A Laboratory Manual Volume II - Cold Spring Harbor Press, Cold Spring Harbor, NY 8.18-8.24 ISBN 0879697164gh
- Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
- Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879695773
Contact
- Sean P Corum
or instead, discuss this protocol.
