Description
Puregene 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 samples to be purified for use as long-term references. To improve availability and address demand more efficiently we implemented an in-house manufacturing solution. This leads to product name and catalogue number changes. This Change will not reflect on the quality and performance of the products. The legacy products will be discontinued on 28.02.2022 and be available only until stocks last. Please use the table in Performance to find the product you need.
The purity of DNA has a significant effect on the accuracy of results obtained in downstream applications. Sensitive downstream applications, such as PCR, demand use of DNA of the highest quality and molecular weight for reliable results. Puregene Kits remove contaminants and enzyme inhibitors, enabling purification of highly stable DNA well-suited for archiving. Purified DNA is of high quality demonstrated by molecular weights of 100–200 kb (see figures "Archive-quality DNA", “ High-molecular-weight DNA from tissue ” and “ High-molecular-weight DNA from mouse tails ”) and performs well in sensitive downstream applications including PCR and restriction digestion (see figure "Efficient restriction endonuclease digestion"). Catalog number changes in the Puregene line Existing Name Cat. no. New name New cat. no Gentra Puregene Blood Kit (120 ml) 158467 Puregene Blood Kit (120 ml) 158023 Gentra Puregene Blood Kit Plus (1000 ml) 158489 Puregene Blood Kit (1000 ml) 158026 Gentra Puregene Cell Kit (8 x 10 8) 158767 Puregene Cell Kit (8 x 10 8) 158043 Gentra Puregene Cell Kit Plus (6.7 x 10 9) 158788 Puregene Cell Kit (6.7 x 10 9) 158046 Gentra Puregene Tissue Kit (4 g) 158667 Puregene Tissue Kit (4 g) 158063 Gentra Puregene Tissue Kit (33 g) 158689 Puregene Tissue Kit (33 g) 158066 Gentra Puregene Blood Kit (1000 ml) 158389 Alternative: Puregene Blood Kit (1000 ml) 158026
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 A 260 /A 280 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.
