MagNA Pure LC Total Nucleic Acid Isolation Kit
Kit for the isolation of total nucleic acid (viral DNA and RNA) from up to 200 µL of mammalian serum, plasma, and whole blood using the MagNA Pure LC Instruments.
For general laboratory use.
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- Use a robot to isolate high-quality viral DNA and RNA for very sensitive downstream PCR.
- Experience superb handsfree reproducibility and sensitivity, difficult to achieve using manual methods.
- Obtain scalable total nucleic acid yields and qPCR results that are a reflection of the amount of starting sample material purified.
- Wash Buffer I, 2 x 100 ml
- Wash Buffer II, 100 ml
- Wash Buffer III, 2 x 100 ml
- Lysis/Binding Buffer, 100 ml
- Proteinase K, 6 glass vials with lyophilizate
- Magnetic Glass Particles (MGPs), 6 vials with 6 ml MGP Suspension each
- Elution Buffer, 100 ml
For general laboratory use.
The MagNA Pure LC Total Nucleic Acid Isolation Kit is designed to isolate high-purity total nucleic acid (e.g., viral DNA and RNA) from mammalian serum, plasma, or whole blood using the MagNA Pure LC Instrument. The purified total nucleic acid can be used in PCR on the LightCycler® Carousel-Based Systems, the LightCycler® 480 System, or standard thermal block cyclers.
Number of isolations/sample type:
192 isolations (6×32) from
- 50 to 200 μl mammalian serum or plasma, or
- 50 to 100 μl mammalian whole blood
For details, see - Instructions for Use
To isolate total nucleic acid from larger-volume samples (up to 1 ml), choose the MagNA Pure LC Total Nucleic Acid Isolation Kit - Large Volume
The isolation procedure is based on magnetic-bead technology. The samples are lysed by incubation with a special buffer containing a chaotropic salt and Proteinase K. Magnetic Glass Particles (MGPs) are added and total nucleic acids contained in the sample are bound to their surface. Unbound substances are removed by several washing steps, then the purified total nucleic acid is eluted with a low-salt buffer.
The principle steps of a MagNA Pure LC total nucleic acid isolation procedure are:
- The sample material is placed into the wells of the Sample Cartridge.
- Lysis/Binding Buffer is added to the sample, resulting in complete cell lysis and release of nucleic acids. Nucleases are denatured.
- Proteinase K is added to the samples and proteins are digested.
- Nucleic acids bind to the silica surface of the added MGPs due to the chaotropic salt conditions, isopropanol, and the high ionic strength of the Lysis/Binding Buffer.
- MGPs with bound nucleic acids are magnetically separated from the residual lysed sample.
- MGPs with bound nucleic acids are washed repeatedly with Wash Buffer to remove unbound substances like proteins (nucleases), cell membranes, PCR inhibitors such as heparin or hemoglobin, and to reduce the chaotropic salt concentration.
- Again MGPs with bound total nucleic acid are magnetically separated from the Wash Buffer containing residual sample debris.
- The purified nucleic acids are eluted at +70°C from the MGPs in the wells of the Elution Cartridge, whereas the MGPs are retained in the reaction tip and discarded.
The MagNA Pure LC Total Nucleic Acid Isolation Kit is function-tested by isolating viral nucleic acid from Hepatitis A Virus (HAV)-positive and Parvo Virus B19-positive human reference material using the 'Total NA Serum_Plasma_Blood' protocol. Purified viral HAV and a Parvo Virus B19-specific nucleic acid was detected using real-time qPCR and qRT-PCR with the LightCycler® 480 and LightCycler® Carousel-Based Instruments. Kit components are also tested to show the absence of nucleases according to the current quality control procedures.