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The High Pure Viral RNA Kit accomplishes virus lysis by incubation of the sample in a special Binding Buffer. Subsequently nucleic acids bind specifically to the surface of glass fibers in the presence of a chaotropic salt (guanidine-HCl). The binding reaction occurs within seconds due to the disruption of the organized structure of water molecules and the interaction of nucleic acids with the glass fibers surface. Thus, adsorption to the glass fiber fleece is favored. Since the binding process is specific for nucleic acids, the bound nucleic acids are purified from salts, proteins and other impurities by a washing step and are finally eluted in low salt Elution Buffer or PCR grade water. The purified viral RNA is free of intact virus, nucleases, and all cellular components that interfere with RT-PCR and can be applied directly for RT-PCR. 50 μl eluate is sufficient for 8-14 RT-PCR reactions.
Included in the kit is a special Inhibitor Removal Buffer resulting in improved sensitivity and reproducibility of RT-PCR assays performed with nucleic acid templates isolated with this kit. Especially, the use of the Inhibitor Removal Buffer allows even the application of heparinized sample material containing 100 U/ml heparin.

The High Pure Viral RNA Kit

• saves time, because the kit does not require extraction with organic solutions or nucleic acid precipitation and thus can prepare multiple RT-PCR templates in approximately 10 minutes
• accommodates a wide variety of samples, because the same kit can purify viral RNA from several bodily fluids
• minimizes RNA loss, because the kit removes contaminants without precipitation or solvent extraction
• increases lab safety, because the kit minimizes the handling of potentially hazardous samples and does not use hazardous organic solvents

1. Serum or plasma are lysed by incubation with Binding Buffer.
2. Nucleic acids are bound to the glass fibers pre-packed in the High Pure Filter Tube.
3. Bound nucleic acids are washed with a special Inhibitor Removal Buffer to get rid of RT-PCR inhibitory contaminants. It allows even the application of heparinized sample material with >100 U/ml heparin.
4. Washing of bound nucleic acids, purification from salts, proteins and other cellular impurities.
5. Purified nucleic acids are recovered using the Elution Buffer.

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Vial / Bottle	Cap	Label	Function / Description	Content
1	green	Binding Buffer	Contains 4.5 M guanidine-HCl, 50 mM Tris-HCl, 30% Triton X-100 (w/v), pH 6.6 (+25°C).	2 × 25 ml
2		Poly (A)	For binding of RNA	2 mg poly(A) carrier RNA (lyophilizate)
3a	black	Inhibitor Removal Buffer	Contains 5 M guanidine-HCl, 20 mM Tris-HCl, pH 6.6 (+25°C) (final concentration after addition of ethanol)	33 ml, add 20 ml absolute ethanol
3	blue	Wash Buffer	20 mM NaCl, 2 mM Tris-HCl, pH 7.5 (+25°C) (final concentrations after addition of ethanol)	2 × 10 ml, add 40 ml absolute ethanol to each vial
4	colorless	Elution Buffer	Water, PCR grade	30 ml
5		High Pure Filter Tubes	For use of up to 700 μl sample volume.	2 bags with 50 polypropylene tubes with two layers of glass fiber fleece
6		Collection Tubes		8 bags with 50 polypropylene tubes (2 ml).

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Total time	Approx. 20 min
Hands-on time	<10 min

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