Difference: RNAPrep (2 vs. 3)

Revision 32018-10-25 - SimonDAlton

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META TOPICPARENT name="ProtocolList"

Isolation of Total RNA

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  • Add 2 volumes (400ul) of RNA Binding Buffer to each volume of RNA sample and mix well.
  • Add 1 volume (600ul) ETOH (95-100%) to the mixture and mix well.
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  • Transfer the mixture to the Zymo-Spin IIC column in a collection tube and centrifuge at >12,000 x g for 1 min. Discard the flow through.
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  • Transfer half the mixture to the Zymo-Spin IIC column in a collection tube and centrifuge at >12,000 x g for 1 min. Discard the flow through.
  • Load the remaining half and repeat spin, discard.
 
  • During this spin, prepare the DNase 1 cocktail for use. Per reaction:
    • 5ul RNase-Free DNase I
    • 75ul Reaction Buffer
 
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