MagNA Pure LC Total Nucleic Acid Isolation Kit - Large Volume
Kit for the isolation of total nucleic acid (viral DNA and RNA) from up to 1,000 µL of mammalian serum and plasma using the MagNA Pure LC Instruments.
For general laboratory use.
- Use a robot to isolate high-quality viral DNA and RNA from large volumes of up to 1 ml of mammalian serum or plasma for very sensitive downstream PCR.
- Experience handsfree reproducibility and sensitivity, difficult to achieve using manual methods.
- Obtain scalable total nucleic acid yields for qPCR and qRT-PCR 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, 4 x 100 ml
- Proteinase K, 6 glass vials with lyophilizate
- Magnetic Glass Particles (MGPs); 6 vials, each containing 11 ml MGP Suspension
- Elution Buffer, 100 ml
For general laboratory use.
The MagNA Pure LC Total Nucleic Acid Isolation Kit - Large Volume is specifically designed to purify total nucleic acid (e.g., viral DNA and RNA) from large amounts (up to 1 ml) of serum and plasma 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 up to 1 ml of mammalian serum or plasmaFor details, see - Instructions for Use
To isolate total nucleic acid from smaller-volume samples (500 to 200 μl), choose the MagNA Pure LC Total Nucleic Acid Isolation Kit
If your instrument is running software version 2.11 (or lower):
To process serum or plasma samples with the MagNA Pure LC Total Nucleic Acid Isolation Kit – Large Volume, you must install a new protocol for the MagNA Pure LC Software, Version 2.11 (or lower). After software installation, the name "Total NA LV Serum_Plasma" should appear in the protocol selection menu on the "Sample Ordering" screen of the MagNA Pure LC. If you do not already have this protocol, you may order it free of charge (Cat. No. 03 188 884 001). For additional details, contact your local Roche representative.
If your instrument is running software version 3.0 or above:
You do not need to install a new protocol. The protocols included in this version of the software will allow you to process serum or plasma with 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 that contains chaotropic salts and Proteinase K. Magnetic Glass Particles are added and the nucleic acid is bound to their surfaces. Unbound substances are removed by several washing steps, and then the purified nucleic acid is eluted.
The principle steps of a MagNA Pure LC DNA 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 acid binds 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 acid are magnetically separated from the residual lysed sample.
- MGPs with bound nucleic acid 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 nucleic acid are magnetically separated from the Wash Buffer containing residual sample debris.
- The purified nucleic acid is 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.