For purification of archive-quality DNA from whole blood, packed cells, buffy coat, and bone marrow
Archive-quality DNA for long-term storage
Reproducible DNA purification
A complete solution for sample-to-storage purification
Convenient, scalable purification procedure
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.
For 1000 ml blood: RBC Lysis Solution, RNase A Solution, and Reagents
The Gentra Puregene Blood Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Puregene DNA procedure.
The simple Puregene procedure uses a modified salting-out precipitation method for purification of DNA. 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.
Efficient restriction endonuclease digestion.
DNA was purified from whole blood samples using the Gentra Puregene Blood Kit. Purified DNA was digested with 2 units enzyme per μg DNA for 2 hours at 37°C. Restriction digestion reactions were analyzed by agarose gel electrophoresis on a 0.6% agarose gel. M: Markers.
Archive-quality DNA.
DNA was purified from replicate whole blood samples using the Gentra Puregene Blood Kit. Purified DNA was analyzed by agarose gel electrophoresis after storage for 18 years at 4°C. M1: Uncut Lambda DNA; M2: Lambda HindIII marker.
Performance
Archive-quality DNA
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").
Principle
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.
Procedure
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.
Applications
DNA purified using the Gentra Puregene Blood Kit is highly stable and suited for use in a wide range of applications, such as:
DNA archiving
PCR
SNP analysis
Southern blotting
Features
Specifications
Applications
PCR, restriction digest, Southern analysis, SNP analysis
Elution volume
250 µl - 1000 µl
Format
Scalable
Main sample type
Whole blood, Buffy coat, body fluid
Processing
Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
2.5 ml (100 mg/ml; 7000 units/ml, solution)
Unit definition: That amount of enzyme causing the hydrolysis of RNA at a rate such that k (velocity constant) equals unity (Kunitz units) at 25°C and pH 5.0.
