MagNA Pure LC DNA Isolation Kit I
Kit for the isolation of genomic DNA from 20 to 200 µL of mammalian whole blood, or 103 to 106 blood or cultured cells, using the MagNA Pure LC Instruments.
For general laboratory use.
- Use a robot to isolate high-quality genomic DNA for sensitive downstream PCR.
- Experience handsfree reproducibility, difficult to achieve using manual methods.
- Obtain scalable DNA yield and quantitative PCR results that are a reflection of the amount of starting sample material purified.
- Lysis/Binding Buffer, 100 ml
- Proteinase K, 6 glass vials with lyophilizate
- Magnetic Glass Particles (MGPs), 6 vials with MGP suspension
- Wash Buffer I, 2 x 100 ml
- Wash Buffer II, 2 x 60 ml
- Elution Buffer, 100 ml
For general laboratory use.
The MagNA Pure LC DNA Isolation Kit I is designed to isolate highly purified genomic DNA from 20 to 200 µl of mammalian whole blood, or 1 x 103 to 1 x 106 blood or cultured cells. Purified DNA in an elution volume fixed to 100 µl, is ideal for real-time qPCR with LightCycler® 480 and LightCycler® Carousel-Based Instruments, and for conventional PCR in thermal block cyclers.
Note: Please obtain the MagNA Pure LC DNA Isolation Kit I – Lysis/Binding Buffer – Refill when requiring additional Lysis/Binding Buffer (bottle 3) for producing large numbers of DNA-stabilized sample lysates from blood, blood cells, and cultured cells.
Number of isolations/sample type:
- 192 isolations (6 x 32)
- 20 to 200 μl mammalian whole blood
- 1 x 103 - 1 x 106 mammalian blood cells (white blood cells [WBCs] or peripheral blood mononuclear cells [PBMCs], or cultured cells)
The isolation procedure is based on magnetic-bead technology. The samples are lysed by incubation with a special buffer containing 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.
- 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.
- 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 human whole blood and cultured cells using the protocol 'DNA I Blood_Cells_High_Performance' and analyzed with respect to DNA integrity, yield, and purity. Purified DNA was also function tested using real-time PCR with LightCycler® 480 and LightCycler® Carousel-Based Instruments.