MagNA Pure LC DNA Isolation Kit III (Bacteria, Fungi)
Kit for the isolation of bacterial and fungal DNA from up to 100 µL sample volumes of a wide variety of sample types using the MagNA Pure LC Instruments.
For general laboratory use.
- Obtain the best possible reproducible results.
- Easily extract high-quality bacterial and fungal DNA using protocols provided for a wide variety of difficult sample materials.
- Quickly isolate bacterial and fungal DNA for the most sensitive quantitative PCR assays.
- 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
- Bacteria Lysis Buffer, 2 x 30 ml
- Proteinase K, 6 glass vials with lyophilizate
For general laboratory use.
The MagNA Pure LC DNA Isolation Kit III (Bacteria, Fungi) is designed to isolate high-purity bacterial and fungal DNA from various research sample materials using the MagNA Pure LC Instrument. The purified DNA can be used in PCR on the
Number of isolations/sample type:
192 isolations (6 x 32) from 50 to 100 μl of liquefied sample (e.g., from bronchoalveolar lavage [BAL], blood culture medium, sputum, cerebrospinal fluid [CSF], urine, swabs, stool, bacterial or fungal cultures).
For details, see Instructions for Use
To process the above-mentioned samples with the MagNA Pure LC DNA Isolation Kit III (Bacteria, Fungi), a new protocol for the MagNA Pure LC Software, Version 2.11 (or lower) must be installed.
The name "DNA III Bacteria" 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. 03 262 774 001). 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 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.
The MagNA Pure LC DNA Isolation Kit III (Bacteria, Fungi) is function-tested by purifying bacterial DNA using the standard protocol from urine spiked with defined numbers of E.coli bacteria. The purified DNA was analyzed with respect to DNA integrity, recovery, purity and ability to amplify target sequence with LightCycler® 480 and LightCycler® Carousel-Based Instruments.