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"Name": "Storage Conditions (Working Solution)",
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Solution | Storage |
Proteinase K | -15 to -25°C |
poly(A) carrier RNA | -15 to -25°C |
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"Language": "en",
"Country": "XG",
"Code": "Storage Conditions (Working Solution)"
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"Value": "Store the set at +15 to +25°C.",
"Language": "en",
"Country": "XG",
"Code": "Storage Conditions (Product)"
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Total time | Approx. 20 minutes |
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Hands-on time | <10 minutes |
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"Name": "Principle",
"Value": "The High Pure Viral Nucleic Acid Buffer Set is intended to be used in combination with the High Pure Viral Nucleic Acid Kit. The kit is designed for the purification of viral nucleic acids from serum or plasma. Nucleic acids can be applied in PCR or RT-PCR directly after elution in nuclease-free water. The purification procedure uses filter tubes instead of extraction with organic solvents or nucleic acid precipitation steps.
The samples are lysed by incubation with a special buffer containing Proteinase K and guanidine hydrochloride that releases nucleic acids (Cory, S. et al., 1983). A highly efficient reaction is obtained at elevated temperatures. Subsequently, the liquid is centrifuged through a glass fiber filter that is contained in the kit (Yang, R. et al., 1979). During this process, the nucleic acids are bound specifically to the surface of the glass fiber (Jakobi, R. et al., 1988; Kristensen, T. et al., 1987). Unbound substances are removed by centrifugation. The absorbed nucelic acids are washed and eluted with an aqueous solution.
- The isolation of the analyte from serum or plasma is required. Virus lysis is accomplished by incubating the sample in a special Lysis / Binding Buffer.
- Isolation of the nucleic acids by binding to the glass fibers.
- Washing of bound nucleic acids, purification from salts, proteins and other cellular impurities.
- Elution of the purified NA with Elution Buffer.
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"Language": "en",
"Country": "XG",
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Vial / Bottle | Cap | Label | Function / Description | Content |
---|
1 | green | Lysis/Binding Buffer | Contains 6 M guanidine-HCl, 10 mM urea, 10 mM Tris-HCl, 20% Triton X-100 (v/v), pH 4.4 (+25°C) | |
2 | white | Poly(A) | Lyophilizate for binding of RNA | |
3 | pink | Proteinase K | Lyophilizate for the digestion of proteins | |
4a | 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
|
4 | blue | Wash Buffer | Contains 20 mM NaCl, 2 mM Tris-HCl, pH 7.5 (+25°C) (final concentrations after addition of ethanol) | - 20 ml add 80 ml absolute ethanol
|
5 | colorless | Elution Buffer | Water PCR grade | |
All solutions are clear, except Vial 1 Lysis / Binding Buffer which is clear to slightly turbid, and should not be used when precipitates have formed. Warm the solutions at +15 to +25°C or in a 37°C water bath until the precipitates have dissolved.
The buffers can show a slight yellow color. This will have no impact on the function of the buffer.
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"Language": "en",
"Country": "XG",
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"Value": "This buffer set supplies additional buffers thereby increasing the flexibility of the High Pure Viral Nucleic Acid Kit.",
"Language": "en",
"Country": "XG",
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"Value": "As a prerequisite for RT-PCR or PCR, viral nucleic acids must be isolated from serum, plasma, or cell-culture supernatant.",
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