Daniel:Protocols/EmulsionPCR: Difference between revisions
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>Djacobse (Created page with "=Emulsion PCR= Back to Main Protocol and tips for emulsion PCR. Used for amplification of highly uniform DNA oligos on streptavidin beads (commonly). ==...") |
>Djacobse |
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Protocol and tips for emulsion PCR. Used for amplification of highly uniform DNA oligos on streptavidin beads (commonly). | Protocol and tips for emulsion PCR. Used for amplification of highly uniform DNA oligos on streptavidin beads (commonly). | ||
[[Media:Shao_Emulsion_PCR_Efficient_DNA_Amplification_PLOS_2011.PDF|Shao Emulsion PCR]] | |||
[[Media:Church_Lab_Emulsion_PCR_Protocol.pdf|Church lab emulsion PCR protocol]] | |||
===Reagents=== | |||
<ol> | |||
<li>Emulsions</li> | |||
<ol type="A"> | |||
<li>Church Lab Protocol</li> | |||
<ol type="a"> | |||
<li>4.01 mL Tegosoft [http://www.sigmaaldrich.com/catalog/product/aldrich/d91551?lang=en®ion=US DEC]</li> | |||
<li>1.10 mL [http://www.sigmaaldrich.com/catalog/product/sigma/m5904?lang=en®ion=US light mineral oil]</li> | |||
<li>385 uL [http://glenncorp.com/shop/abil-09/ ABIL WE 09]</li> | |||
<li>Add 960 uL aqueous solution in 5.5 mL oil phase in a 50 mL falcon tube; vortex 5 minutes</li> | |||
</ol> | |||
<li>Shao et al Protocol</li> | |||
<ol type="a"> | |||
<li>4.5% Span 80</li> | |||
<li>0.4% Tween 80</li> | |||
<li>0.05% Triton X-100</li> | |||
<li>Mix 100 uL aqueous, 10 uL at a time, into 200 uL oil phase in a 2 mL cryo-vial continuously stirred at 1500 rpm with a [https://us.vwr.com/store/catalog/product.jsp?catalog_number=58948-377 magnetic microstir bar]</li> | |||
</ol> | |||
<li> | |||
==Protocol== | ==Protocol== |
Revision as of 20:31, 10 April 2017
Emulsion PCR
Protocol and tips for emulsion PCR. Used for amplification of highly uniform DNA oligos on streptavidin beads (commonly).
Church lab emulsion PCR protocol
Reagents
- Emulsions
- Church Lab Protocol
- 4.01 mL Tegosoft DEC
- 1.10 mL light mineral oil
- 385 uL ABIL WE 09
- Add 960 uL aqueous solution in 5.5 mL oil phase in a 50 mL falcon tube; vortex 5 minutes
- Shao et al Protocol
- 4.5% Span 80
- 0.4% Tween 80
- 0.05% Triton X-100
- Mix 100 uL aqueous, 10 uL at a time, into 200 uL oil phase in a 2 mL cryo-vial continuously stirred at 1500 rpm with a magnetic microstir bar
-
Protocol