Gentra Puregene Blood Kit
For purification of archive-quality DNA from whole blood, packed cells, buffy coat, and bone marrow
Gentra Puregene Blood Kits enable purification of high-molecular-weight (100–200 kb) DNA suitable for archiving. The scalable purification procedure gently removes contaminants and inhibitors and allows large-volume samples to be purified for use as long-term references.
An ongoing stability study of archived DNA has shown that high-molecular-weight (100–200 kb) DNA purified using Gentra Puregene Kits can be stored at 4°C for at least 18 years without degradation (see figure "Archive-quality DNA"). The scalable purification procedure allows large sample volumes to be conveniently handled. The combination of large-scale preparation and archive-quality DNA is highly suited for applications such as genetic research, biobanking, and clinical trials that require long-term references from sample sources of limited availability or from samples that must be used anonymously.
High performance in sensitive downstream applications
Purity of DNA has a significant effect on the accuracy of results obtained in downstream applications. Sensitive downstream applications, such as PCR, demand the use of DNA of the highest quality and molecular weight for reliable results. The Gentra Puregene Blood Kit removes contaminants and enzyme inhibitors enabling purification of high-quality DNA. Purified DNA is ready to use and performs well in sensitive downstream applications including PCR and restriction digestion (see figure "Efficient restriction endonuclease digestion").
Cells are lysed with an anionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in hydration solution (1 mM EDTA, 10 mM Tris·Cl pH 7.5). Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2–8°C, –20°C, or –80°C.
The simple Puregene procedure uses a modified salting-out precipitation method for purification of DNA (see flowchart "Puregene DNA procedure"). No mixing or dilution of solutions is necessary, and hands-on time is minimized. The procedure provides convenient stopping points that allow safe, temporary storage of partially purified samples.
DNA purified using the Gentra Puregene Blood Kit is highly stable and suited for use in a wide range of applications, such as:
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