Barrick Lab :: ProceduresTotalAlkalineDigestOfEmbeddedRNALinkages

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---+!! Total Alkaline Digest of Embedded RNA Linkages

---++ Materials

   *   0.2 N Sodium Hydroxide (NaOH)  
      *   Molarity and Normality are related by N = nM For example, a 0.01 M solution of H<sub>2</sub> SO <sub>4</sub> is 0.02 N. 
   *   3 M Sodium Acetate (NaOAc)
   *   100% Ethanol (EtOH)

---++ Procedure

   1 Precipitate PCR DNA with embedded ribose with ethanol.
   1 Resuspend in 0.9 of original PCR volume with 0.2 N NaOH.
   1 Heat at 90&deg;C, for 10 min. 
   1 Add 3 M NaOAc, about 1/9 the volume of the sample,  to neutralize and 2.5 volumes cold ethanol if [[ProtocolsEthanolPrecipitation][Ethanol Precipitation]] is needed for a later step.   
      
      *Modified from Breaker Laboratory Protocols, 2004 Protocols




-- Main.AlvaroRodriguez - 16 Jul 2012

Barrick Lab > ProtocolList > ProceduresTotalAlkalineDigestOfEmbeddedRNALinkages

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TWikiGuest, AlvaroRodriguez

Topic revision: r2 - 2013-08-21 - 23:05:46 - Main.AlvaroRodriguez