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: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&region=US DEC]</li>
<li>1.10 mL [http://www.sigmaaldrich.com/catalog/product/sigma/m5904?lang=en&region=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

Back to Main

Protocol and tips for emulsion PCR. Used for amplification of highly uniform DNA oligos on streptavidin beads (commonly).

Shao Emulsion PCR

Church lab emulsion PCR protocol


Reagents

  1. Emulsions
    1. Church Lab Protocol
      1. 4.01 mL Tegosoft DEC
      2. 1.10 mL light mineral oil
      3. 385 uL ABIL WE 09
      4. Add 960 uL aqueous solution in 5.5 mL oil phase in a 50 mL falcon tube; vortex 5 minutes
    2. Shao et al Protocol
      1. 4.5% Span 80
      2. 0.4% Tween 80
      3. 0.05% Triton X-100
      4. 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
    3. Protocol