This protocol utilizes the powerful guanidine isothiocyanate–phenol:chloroform extraction method which allows the rapid isolation of total RNA including small RNAs. The conditions for extraction enable the partitioning of proteins and DNA into the organic layer of the biphasic solution interface, while retaining RNA in the upper aqueous layer. The aqueous phase is removed to a second tube, and RNA is precipitated with an equal volume of isopropanol. High yields of pure, undegraded total RNA can be recovered from even small quantities of tissue or cells. Large numbers of samples may be performed simultaneously, because of the simplicity of the technique.
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