Daniel:Protocols/EmulsionPCR: Difference between revisions
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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:Shao_Emulsion_PCR_Efficient_DNA_Amplification_PLOS_2011.PDF|Shao Emulsion PCR (2011)]] | ||
[[Media:Church_Lab_Emulsion_PCR_Protocol.pdf|Church lab emulsion PCR protocol]] | [[Media:Church_Lab_Emulsion_PCR_Protocol.pdf|Church lab emulsion PCR protocol (2007)]] | ||
[[Media:Murgha_Methods_For_ssOligo_Libraries_PLOS_One_2014.PDF|Murgha et al protocol (2014)]] | |||
===Reagents=== | ===Reagents=== | ||
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<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> | <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> | </ol> | ||
<li>Murgha et al | <li>Murgha et al</li> | ||
<ol type="a"> | |||
<li>4% [http://glenncorp.com/shop/abil-09/ ABIL WE 09]</li> | |||
<li>0.05% TritonX-100</li> | |||
<li>Mineral oil (solvent)</li> | |||
<li>Add aqueous to oil and stir at 1000 rpm at 4C; stir additional 15 minutes after mixing</li> | |||
</ol></ol> | |||
<li>PCR protocols</li> | |||
<ol type="A"> | |||
<li>Church Lab Protocol</li> | |||
</ol> | |||
==Protocol== | ==Protocol== |
Revision as of 20:44, 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 (2007)
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
- Murgha et al
- 4% ABIL WE 09
- 0.05% TritonX-100
- Mineral oil (solvent)
- Add aqueous to oil and stir at 1000 rpm at 4C; stir additional 15 minutes after mixing
- PCR protocols
- Church Lab Protocol