How To Make A MEGA-plate: Difference between revisions
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* [[Image:Polycarbonate sheet 60cm X 80cm]] | * [[Image:Polycarbonate sheet 60cm X 80cm]] | ||
* Plexiglas sheet 30cm X 60cm | * Plexiglas sheet 30cm X 60cm | ||
* [[Image:Dichloromethane.jpg]] | * Dichloromethane | ||
[[Image:Dichloromethane.jpg]] | |||
* Syringe | * Syringe | ||
* Needle | * Needle |
Revision as of 05:29, 12 January 2017
Overview
Replace this sentence with a brief description of the protocol and its goal.
The goal is hoe to build a MEGA-plate.
Materials
For a plate of 60cm on 80cm:
- File:Polycarbonate sheet 60cm X 80cm
- Plexiglas sheet 30cm X 60cm
- Dichloromethane
- Syringe
- Needle
- 9 X 2L Erlenmeyer flask
- File: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
- Goldbeter, A and Koshland, DE (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
- 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 0879697164
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.