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illustra blood genomicPrep Mini Spin Kit- For the rapid and reproducible isolation of high-quality genomic DNA from whole blood, buffy coat, bone marrow, and nucleated red blood cells.
- High-quality genomic DNA with a greater proportion of higher molecular weight DNA and less shearing.
- Demonstrated application to various types of whole blood including human, horse, rabbit, rat and mouse.
- Increased ease of use with reduced pipetting volume changes, one centrifugation setting, and color-coded kit components.
- Validated in several downstream applications including real-time PCR, endpoint PCR, multiplex PCR, and restriction enzyme digests.
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illustra blood genomicPrep Mini Spin Kit
Technical Information
Real-time PCR amplification from genomic DNA extracted from K3 EDTA-treated whole blood using the illustra blood genomicPrep Mini Spin Kit. The eluted product (3 μl ) and various amounts (0.1, 1, 10, and 100 ng) of control DNA were amplified using chromosome-specific primers in a real-time PCR assay on an ABI 7900. Replicates were n = 3.
The purification process involves minimal shearing resulting in the production of 5 to 10 μg of good quality, intact genomic DNA from a 200 μl sample. Input sample volumes can vary from 50 μl to 1 ml. The method uses a simple genomic DNA purification protocol that uses chaotropic agents to extract DNA from blood cells, denature protein components, and promote the selective binding of DNA to a column-based, novel silica-membrane. The elution volume can be modified as necessary to obtain the appropriate genomic DNA concentration for most downstream molecular biological applications. The kit contains spin columns prepacked with a novel silica membrane, lyophilized proteinase K powder, lysis solution, wash and elution buffers, microcentrifuge collection tubes, a full protocol booklet, and a detachable, quick reference protocol card.
Digestion of purified genomic DNA with restriction enzymes. Purified genomic DNA from the blood genomicPrep Mini Spin Kit was digested with AluI, EcoRI, HindIII, and BamHI restriction enzymes.
Kit compatibility across multiple anticoagulants. Isolation of genomic DNA was carried out from 200 μl of human (Hu) whole blood treated with K3-EDTA (EDTA), sodium citrate (Cit) and sodium heparin (Hep). The plot shows mean values for yield and purity with standard deviations.
| TECHNICAL SPECIFICATIONS |
| Yield | 4 to 6 μg/200 μl whole blood |
| Purity | > 1.7 to 1.6 |
| Total time of prep* | less than 20 min |
| Hands-on time* | less than 5 min |
| Size | > 20 kbp |
| *including lysis, purification & desalting
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References
- Sambrook, J., Fritsch, E.F and Maniatis, T. Molecular cloning: A laboratory Manual, Cold Spring Harbor laboratory, 2nd eds., (1989).
- Vogelstein, B. and Gillespie, D. Proc. Natl. Acad. Sci. USA 76, 615 (1979).
- Marko, M.A., Chipperfield, R. and Birnboim, H.C. Anal. Biochem. 121, 382 (1982).
- Voo, K.S. and Jacobsen B.M., Biotechniques 24, 240-243 (1998).
- Hilz, H. et al. Eur. J. Biochem. 56, 103-108 (1975).
- Ausubel, F.M. et al., eds., Current Protocols in Molecular Biology 1, 1.68 (1991).
- Melzak, K. A. et al., J. Coll. Interf. Sci. 181, 635-644 (1996).
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Data file
 and various amounts (0.1, 1, 10, and 100 ng) of control DNA were amplified using chromosome-specific primers in a real-time PCR assay on an ABI 7900. Replicates were n = 3.'))
)
 whole blood treated with K3-EDTA (EDTA), sodium citrate (Cit) and sodium heparin (Hep). The plot shows mean values for yield and purity with standard deviations.'))
