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 H₂ SO ₄ is 0.02 N.
• 3 M Sodium Acetate (NaOAc)
• 100% Ethanol (EtOH)

Procedure

1. Precipitate PCR DNA with embedded ribose with ethanol.
2. Resuspend in 0.9 of original PCR volume with 0.2 N NaOH.
3. Heat at 90°C, for 10 min.
4. Add 3 M NaOAc, about 1/9 the volume of the sample, to neutralize and 2.5 volumes cold ethanol if Ethanol Precipitation is needed for a later step.

*Modified from Breaker Laboratory Protocols, 2004 Protocols

-- Main.AlvaroRodriguez - 16 Jul 2012