MagNA Pure LC DNA Isolation Kit II (Tissue)
Kit for the isolation of genomic DNA from one to ten milligrams of mammalian tissues including fresh-frozen tissue, and formalin-fixed, paraffin-embedded (FFPE) tissue sections using the MagNA Pure LC Instruments.
For general laboratory use.
- Easily isolate high-quality DNA from a broad range of even the most difficult tissue types (including paraffin sections) using the optimized external tissue lysis.
- Quickly obtain DNA of high integrity and purity suitable for highly sensitive PCR.
- Achieve excellent reproducible DNA yield for exquisite quantitative applications.
- Wash Buffer I, 2 x 100 ml
- Wash Buffer II, 2 x 60 ml
- Wash Buffer III, 100 ml
- Lysis/Binding Buffer, 100 ml
- Magnetic Glass Particles (MGPs), 6 vials with MGP suspension
- Elution Buffer, 100 ml
- Tissue Lysis Buffer, 100 ml
- Proteinase K, 6 glass vials with lyophilizate
For general laboratory use.
The MagNA Pure LC DNA Isolation Kit II (Tissue) is designed to isolate high-purity DNA from mammalian tissues 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.
Number of isolations/sample type:
192 isolations (6 x 32) from:
- 1 to 10 mg fresh or frozen mammalian tissue, or
- 5 to 10 μm sections of formalin-fixed, paraffin-embedded (FFPE) mammalian tissue
For details, see - Instructions for Use
To process tissue samples with the MagNA Pure LC DNA Isolation Kit II (Tissue), a new protocol for the MagNA Pure LC Software, Version 2.11 (or lower) must be installed.
The name "DNA II Tissue" should appear in the protocol selection of the "Sample Ordering" screen of the MagNA Pure LC Software. If not previously installed, order the protocol free of charge (Cat. No. 03 190 820 001). If running software version 3.0 or above, no extra protocol installation is required. For additional details, contact your local Roche representative.
The isolation procedure is based on magnetic-bead technology. The samples are lysed by incubation with a special buffer that contains chemotropic 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 is isolated from mouse tissue using the standard protocol and analyzed with respect to integrity, yield, purity and ability to amplify in the LightCycler® Carousel-Based Instrument.