Principle / Procedure

The extraction of nucleic acids from biological matrices like serum, plasma and other cell-free solutions becomes more and more important. Typically available spin column devices for the purification of viral nucleic acids can be used with up to 200 μL of plasma or serum in one loading step. If larger amounts (e.g., pooled samples) are to be processed, standard spin columns have to be loaded successively, thus increasing the risk of cross-contamination.
For this reason MACHEREY-NAGEL has developed a column for the processing of larger volumes. The funnel column of the NucleoSpin® RNA Virus F kit allows handling of up to 1 mL serum or plasma with only one loading step. Due to the small diameter of the membrane the elution volume is still between 50–100 μL (low-salt buffer or water), resulting in a highly concentrated sample directly suitable for any amplification process. The funnel column can be centrifuged in a 50 mL polyethylene tube (included in the kit) and can be closed separately. Elution tubes are fitted directly onto the funnel column. Cross contamination is eliminated because all steps are performed in a closed system. Eight or twelve samples (depending on the centrifuge rotor used) can be processed in 30 minutes.

Features

• Up to 1 mL of cell-free biological fluids can be processed – suitable for large sample volumes
• Ideal for pooled samples
• Patented technology of NucleoSpin® Funnel Columns
• Closed system preventing cross-contamination for highest safety
• Small elution volume of 50–100 μL leads to concentrated RNA and highest sensitivity

Technical specifications

Applications**

• Viral RNA from up to 1 mL serum, plasma, cell-free biological fluids
• HCV, HBV, HAV, HIV, HSV, HPV, VZV, EBV, parvovirus B19, H5N1, H1N1
• Support protocol for parallel isolation of viral RNA and DNA*
• Typical downstream applications: RT-PCR, enzymatic reactions

* For parallel isolation of viral RNA and DNA use of Proteinase K required (not included in the kit), see art. # 740506
** Kits to be used for research purposes only