MagNA Pure LC DNA Isolation Kit - Large Volume
Kit for the isolation of genomic DNA from 20 µL up to 1 mL of mammalian whole blood, blood cells, or cultured cells using the MagNA Pure LC Instruments.
For general laboratory use.
- Quickly isolate genomic DNA of very high quality and integrity using a robot.
- Reproducibly obtain genomic DNA from up to 32 samples, each with up to 1 ml of whole blood, in 30 to 177 minutes.
- Flexibly process different sample volumes choosing from four different purification protocols.
- Wash Buffer I, 3 x 100 ml
- Wash Buffer II, 3 x 60 ml
- Wash Buffer III, 2 x 100 ml
- Lysis/Binding Buffer, 100 ml
- Magnetic Glass Particles (MGPs) Suspension, 6 vials with MGP suspension
- Elution Buffer, 100 ml
- Proteinase K Buffer II, 100 ml
- Proteinase K, 3 glass vials with lyophilizate
For general laboratory use.
The MagNA Pure LC DNA Isolation Kit – Large Volume is designed to isolate high-purity genomic DNA from mammalian whole blood, blood cells, or cultured cells using the MagNA Pure LC Instrument. The purified DNA can be used in PCR on the LightCycler ® Carousel-Based Systems, the LightCycler ® 480 System, or standard thermal block cyclers.
The MagNA Pure LC DNA Isolation Kit - Large Volume is used for automated isolation of genomic DNA from up to 5 x 106 cells. The kit is designed for:
- 96 isolations from 1 ml mammalian whole blood (3 runs with 32 samples)
- 192 isolations from 300 to 500 µl mammalian whole blood (6 runs with 32 samples)
- 288 isolations from 20 to 200 µl mammalian whole blood (9 runs with 32 samples each)
- 192 isolations from mammalian blood cells or cultured cells
To perform DNA isolations with the MagNA Pure LC DNA Isolation Kit – Large Volume, new protocols for the MagNA Pure LC Software Version 2.11 (or lower) must be installed. If not previously installed, order the protocols (Cat. No. 03 328 538 001) on floppy disc free of charge. If running software version 3.0 or above, no extra protocol installation is required. For additional details, contact your Roche representative.
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 DNA is bound to their surfaces. Unbound substances are removed by several washing steps, then the purified DNA 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.
- Proteinase K is added to the samples and proteins are digested.
- Lysis/Binding Buffer is added to the sample, resulting in complete cell lysis and release of nucleic acids. Nucleases are denatured.
- DNA 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 DNA are magnetically separated from the residual lysed sample.
- MGPs with bound DNA 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 DNA acid are magnetically separated from the Wash Buffer containing residual sample debris.
- The purified DNA 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.
DNA was isolated from one milliliter of human blood and analyzed with respect to integrity, yield, purity, and ability to be amplified on LightCycler® Carousel-Based Instruments.