<noautolink> ---+!! 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°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
This topic: Lab
>
WebHome
>
ProtocolList
>
ProceduresTotalAlkalineDigestOfEmbeddedRNALinkages
Topic revision: r2 - 2013-08-21 - AlvaroRodriguez