Description

MagExtractor -Viral RNA- provides a simple and reliable method for the rapid purification of viral RNA from serum or plasma specimens using magnetic silica beads. This kit is based on the property that RNA can be absorbed onto a silica surface in the presence of chaotropic agents (1),(2) and an RNA-binding accelerator. The purified viral RNA can be used directly for RT-PCR experiments.

Features

  • This kit is suitable for the high-throughput extraction of viral RNA from serum or plasma specimens using
    magnetic silica beads.
  • This kit does not contain hazardous substances such as phenol or chloroform.
  • No ethanol is used in the washing steps.

Applications

  • Purification of viral RNA from serum or plasma specimens.

Storage condition

Store at 4ºC

Components

Lysis & Binding Solution* 77 ml
Washing Solution I 154 ml
Washing Solution II 200 ml
Elution Solution 10 ml
Magnetic Beads 8 ml

*1/100 volume of 2-mercaptoethanol must be added prior to use.

Principle

The selectivity of extracted nucleic acids can be changed by optimization of the binding and washing solutions. MagExtractor -Genome- (Code No. NPK-101) extracts genomic DNA from various specimens (e.g. whole blood, cultured cells or animal tissues etc.). MagExtractor -RNA- (Code No. NPK-201) extracts total RNA from various specimens (e.g. cultured cells or animal tissues). MagExtractor -Plasmid- (Code No. NPK-301) extracts plasmids from E. coli cells, MagExtractor -Viral RNA- (Code No. NPK-401) is a kit for extracting viral RNA from serum or plasma specimens. MagExtractor -Plant Genome- (Code No. NPK-501) is a kit for extracting genomic DNA from various plant specimens (e.g., leaf, cultured cells, etc.). MagExtractor-PCR & Gel Clean up- (Code No. NPK-601) extracts DNA fragments from a PCR solution, enzyme solution, or agarose gel slices.

Application data

Example 1. Purification of viral RNA from HCV (hepatitis C virus) positive serum and detection of the viral RNA by RT-PCR

HCV RNA was purified from serially diluted HCV-positive serum specimens (100 µl; 102-104 copies) using various purification kits and detected by RT-PCR using 1/40 volume of purified RNA solution. As shown in Fig.1, HCV RNA could be detected down to 103 copies by use of MagExtractor -Viral RNA-.

Fig. 1. Detection of HCV by RT-PCR

References

  • B. Vogelstein and D. Gillespie, Proc. Natl. Acad. Sci. USA. 76: 615-619 (1979)
  • R. Boom, C. J. A. Sol, M. M. M.Salimans, C. L. Wertheim-van Dillen, P. M. E. Dillen and J. van der Noordaa, J. Clin. Microbiol., 28: 495-503 (1990)