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
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Gentra Puregene Blood Kit (30 ml)

Cat. No. / ID: 158445

For 30 ml blood: RBC Lysis Solution, RNase A Solution, and Reagents
Kit
Gentra Puregene Kit
Puregene Kit
Sample type
Blood
Buccal Cells
Yeast/Bacteria
For
30 ml
1000 ml
This product will be discontinued as of October 31, 2022 or until stocks last. We recommend 158023.
Puregene Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

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.

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.

 

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. 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 Purgene 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 108) 158767 Puregene Cell Kit (8 x 108) 158043
Gentra Puregene Cell Kit Plus (6.7 x 109) 158788 Puregene Cell Kit (6.7 x 109) 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

 

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 (4)
This protocol is designed for purification of DNA from 0.5 or 1 ml samples of Gram-positive bacteria culture medium using the Gentra Puregene Yeast/Bact. Kit.
This protocol is designed for purification of DNA from 300 μl samples of whole blood using the Gentra Puregene Yeast/Bact. Kit.
This protocol is designed for purification of DNA from 5 ml samples of Gram-positive bacteria culture medium using the Gentra Puregene Yeast/Bact. Kit.
This protocol is designed for purification of DNA from 1 g samples of soil using the Gentra Puregene Yeast/Bact. Kit.
Kit Handbooks (1)
For purification of archive-quality DNA from human whole blood, bone marrow, buffy coat, buccal cells, body fluids, cultured cells, tissue, mouse tail, yeast, bacteria
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Instrument Technical Documents (1)

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

FAQ

Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728