Gentra Puregene Kits

For purification of archive-quality DNA from a wide variety of sample types

Features

  • High-molecular-weight DNA in the range of 100–200 kb
  • Archive-quality DNA isolation for long-term storage
  • Reproducible DNA purification
  • A complete solution for sample-to-storage purification
  • Convenient, scalable purification procedure

Product Details

Gentra 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.

Performance

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. Gentra 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").

See figures

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.

See figures

Applications

DNA purified using Gentra Puregene Kits is highly stable and suited for use in a wide range of applications, such as:

  • DNA archiving
  • PCR
  • SNP analysis
  • Southern blotting

Comparison of Gentra Puregene Kits

Features Gentra Puregene Tissue Kit Gentra Puregene Buccal Cell Kit Gentra Puregene Cell Kit Gentra Puregene Blood Kit Gentra Puregene Mouse Tail Kit Gentra Puregene Yeast/Bact. Kit
Applications PCR, restriction digest, Southern analysis, SNP analysis PCR, restriction digest, Southern analysis, SNP analysis PCR, restriction digest, Southern analysis, SNP analysis PCR, restriction digest, Southern analysis, SNP analysis PCR, restriction digest, Southern analysis, SNP analysis PCR, restriction digest, Southern analysis, SNP analysis
Elution volume 50–250 µl (varies) 20 µl (varies) 50–250 µl (varies) 250 µl – 1000 µl 50 µl (varies) 50 µl (varies)
Format Scalable Scalable Scalable Scalable Scalable Scalable
Main sample type Tissue samples, fixed and paraffin-embedded tissue Buccal brush, mouth wash Cultured cells Whole blood, buffy coat, body fluid Mouse tail Gram+ and Gram- bacteria, yeast
Processing Manual Manual Manual Manual Manual Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Genomic DNA Genomic DNA Genomic DNA Genomic DNA Genomic DNA Bacterial DNA / yeast DNA
Sample amount 5–100 mg 1 swab / 10 ml 1 x 106 – 2.2 x 107 300 µl – 10 ml / 1 x 107 / 50 µl – 1000 µl 5–10 mg 0.5–1.5 x 109 cells / 1–2 x 108 cells
Technology Modified salting-out precipitation method Modified salting-out precipitation method Modified salting-out precipitation method Modified salting-out precipitation method Modified salting-out precipitation method Modified salting-out precipitation method
Time per run or per prep 25–60 minutes (+ cell lysis + DNA rehydration) 25–60 minutes (+ cell lysis + DNA rehydration) 25–60 minutes (+ DNA rehydration) 25–60 minutes (+ DNA rehydration) 25–60 minutes (+ cell lysis + DNA rehydration) 25–60 minutes (+ DNA rehydration)
Yield Varies 1 µg/swab / 46 µg/10 ml mouth wash 7 µg / 1 million cells 35 µg / ml whole blood 10–75 µg 17–50 µg/ml culture / 4.5 µg/ml culture

Supporting data and figures

Resources

Supplementary Protocols (0)
Kit Handbooks (0)
Safety Data Sheets (0)
Instrument Technical Documents (0)

Publications

Methylenetetrahydrofolate reductase polymorphisms and therapy response in pediatric acute lymphoblastic leukemia.
Aplenc R; Thompson J; Han P; La M; Zhao H; Lange B; Rebbeck T;
Cancer Res; 2005; 65 (6):2482-7 2005 Mar 15 PMID:15781665
High-density single nucleotide polymorphism array defines novel stage and location-dependent allelic imbalances in human bladder tumors.
Koed K; Wiuf C; Christensen LL; Wikman FP; Zieger K; Møller K; von der Maase H; Orntoft TF;
Cancer Res; 2005; 65 (1):34-45 2005 Jan 1 PMID:15665277
Hematopoietic cells and osteoblasts are derived from a common marrow progenitor after bone marrow transplantation.
Dominici M; Pritchard C; Garlits JE; Hofmann TJ; Persons DA; Horwitz EM;
Proc Natl Acad Sci U S A; 2004; 101 (32):11761-6 2004 Jul 28 PMID:15282377
A novel technique based on a PNA hybridization probe and FRET principle for quantification of mutant genotype in fibrous dysplasia/McCune-Albright syndrome.
Karadag A; Riminucci M; Bianco P; Cherman N; Kuznetsov SA; Nguyen N; Collins MT; Robey PG; Fisher LW;
Nucleic Acids Res; 2004; 32 (7):e63 2004 Apr 19 PMID:15096559
High incidence of somatic mutations in the AML1/RUNX1 gene in myelodysplastic syndrome and low blast percentage myeloid leukemia with myelodysplasia.
Harada H; Harada Y; Niimi H; Kyo T; Kimura A; Inaba T;
Blood; 2003; 103 (6):2316-24 2003 Nov 13 PMID:14615365