Daniel:Protocols/EmulsionPCR: Difference between revisions

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>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: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

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

Shao Emulsion PCR (2011)

Church lab emulsion PCR protocol (2007)

Murgha et al protocol (2014)

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. Murgha et al
      1. 4% ABIL WE 09
      2. 0.05% TritonX-100
      3. Mineral oil (solvent)
      4. Add aqueous to oil and stir at 1000 rpm at 4C; stir additional 15 minutes after mixing
  2. PCR protocols
    1. Church Lab Protocol

    Protocol