Bio-Rad PureZOL™ RNA Isolation Reagent - Manual (Page 3)

Section 1
Introduction
PureZOL RNA isolation reagent is intended for the extraction of
total RNA from animal and plant tissues, cultured mammalian
cells, and bacterial and yeast cells in under 1 hour. PureZOL can
also be used for the simultaneous extraction of RNA, DNA, and
proteins from various samples. This reagent allows processing of
small amounts of starting material (50 cells or 5 mg of tissue), and
can be scaled up to process large amounts of starting material.
PureZOL RNA isolation reagent is a monophasic solution of
phenol and guanidine isothiocynate, an improved version of the
single-step isolation of total RNA developed by Chomczynski and
Sacchi (1987). The reagent facilitates immediate and effective
inhibition of RNase activity, while lysing cells and eliminating other
cellular components. Following the addition of chloroform and
subsequent centrifugation, the homogenate separates into an
aqueous phase, an interphase and an organic phase. RNA is
recovered in the aqueous phase with the addition of isopropyl
alcohol, and is subsequently washed in ethanol and solubilized in
RNase-free water.
Total RNA isolated from this procedure is generally DNA- and
protein-free and can be used for northern blot analysis,
in vitro
translation, poly (A) + selection, RNase protection assays, RT-PCR,
and molecular cloning. For RT-PCR analysis, DNase treatment may
be necessary for optimal results. Subsequent cleanup of RNA using
an Aurum™ total RNA mini kit (732-6820) is recommended to
remove any phenol and other contaminants that may have
coprecipitated with the RNA (see section 8 for ordering information).
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