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Solution | Storage |
PolyA carrier RNA solution | -15 to -25°C |
Proteinase K | -15 to -25°C |
",
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"Code": "Storage Conditions (Working Solution)",
"Name": "Storage Conditions (Working Solution)"
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"Value": "Store the kit at +15 to +25°C.",
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"Value": "For isolation of viral nucleic acids for PCR and RT-PCR",
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"Value": "Isolation of the analyte from serum, plasma or whole blood is required as a pre-requisite for the analysis of viral nucleic acids by the polymerase chain reaction PCR or RT-PCR.
Virus lysis is accomplished by incubation of the sample in a special Lysis/Binding buffer in the presence of Proteinase K. Subsequently, nucleic acids bind specifically to the surface of glass fibers in the presence of a chaotropic salt. 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 surface of the glass fibers, thereby promoting adsorption to the glass fiber fleece. 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 eluted in low salt buffer or water.
- Serum, plasma, or whole blood are lysed by incubation with binding buffer and Proteinase K.
- Nucleic acids are bound to the glass fibers pre-packed in the High Pure Filter Tube.
- Bound nucleic acids are washed with a special Inhibitor Removal Buffer to get rid of PCR inhibitory contaminants.
- Washing of bound nucleic acids, purification from salts, proteins and other cellular impurities.
- Purified nucleic acids are recovered using the Elution Buffer.
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Total time | Approx. 25 min |
---|
Hands-on time | 10 min |
---|
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Vial / Bottle | Cap | Label | Function / Description | Content |
---|
1 | green | Binding Buffer | Contains 6 M guanidine-HCl, 10 mM Tris-HCl, 10 mM urea, 20% -Triton X-100 w/v, pH 4.4 +25°C | |
2 | | PolyA | Lyophilizate poly(A) carrier RNA 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 | - 10 ml, add 40 ml absolute ethanol each case
|
5 | colorless | Elution Buffer | Water, PCR Grade | |
6 | | High Pure Extender Assembly | | - 8 bags, 5 pieces each in a single zip pack
|
7 | | Collection Tubes | | - 2 bags with 50 polypropylene tubes 2 ml
|
All solutions are clear except Vial 1, 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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"Value": "The High Pure Viral Nucleic Acid Large Volume Kit is designed for the purification of viral nucleic acids from up to 2.5 mL of mammalian serum, plasma or whole blood.
When using whole blood, total nucleic acids are purified, including viral nucleic acids. The purified viral nucleic acids are applied in PCR or RT-PCR directly after elution in nuclease-free water.",
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