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
Gentra Puregene Blood Kit (30 ml)

Cat. No. / ID: 158445

For 30 ml blood: RBC Lysis Solution, RNase A Solution, and Reagents
$131.00
Kit
Gentra Puregene Kit
Gentra Puregene Kit Plus
Sample type
Blood
Cells
Tissue
Buccal Cells
Yeast/Bacteria
Mouse Tail
For
30 ml
120 ml
1000 ml
Add to cart
Gentra Puregene Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

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 (42)
This protocol is designed for purification of DNA from 0.5-2 mg samples of Eimeria oocysts using Gentra Puregene Cell Kits.
This protocol provides information about scaling of reagents required for purification of DNA from buffy coat prepared from 0.3-15 ml samples of whole blood using the Gentra Puregene Blood Kit.
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 provides information about scaling of reagents required for purification of DNA from 5-60 mg mouse tail tissue using the Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from blood smears on a slide using Gentra Puregene Cell Kits.
This protocol is designed for rapid purification of DNA from 1-2 x 106, 3-5 x 106, or 6-9 x 106 cells using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 2 ml samples of umbilical cord blood using the Gentra Puregene Tissue Kit or Gentra Puregene Blood Kit.
This protocol is designed for purification of DNA from 100 Drosophila melanogaster using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 5-20 mg samples of marine invertebrate tissue using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 50 μl or 1 ml samples of clotted whole blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 5 buccal brushes using the Gentra Puregene Buccal Cell Kit.
This protocol is designed for purification of DNA from 100-150 mg dried or 200-400 mg fresh leaf tissue using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 0.5 ml or 1 ml Gram-negative bacteria culture medium using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 5-7 ml clotted whole blood using Clotspin Baskets and the Gentra Puregene Blood Kit.
This protocol provides information about scaling of reagents required for purification of DNA from 1 x 106 - 3 x 107 fixed cells using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 600-1200 mg fresh or frozen leaf tissue or 300-450 mg dried leaf tissue using the Gentra Puregene Cell 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 300 μl samples of whole blood using the Gentra Puregene Blood Kit.
This protocol is designed for purification of DNA from 300-700 (300-700 mg) Drosophila melanogaster using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 5-10 mg or 10-20 mg samples of solid tissue fixed in ethanol or formalin using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 1 g samples of soil using the Gentra Puregene Yeast/Bact. Kit.
This protocol provides information about scaling of reagents required for purification of DNA from 0.5-20 mg paraffin-embedded tissue using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 10-20 mg samples of fish tissue using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 25 ml Gram-negative bacteria culture medium using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 1-2 ml samples of Chlamydomonas using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 5-10 mg or 10-20 mg samples of fungal tissue using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol provides information about scaling of reagents required for purification of DNA from 100 to 5 x 108 cultured cells using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 50 μl samples of CSF using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 40-90 mg (50-75 μl), 80-180 mg (100-150 μl), or 20 g samples of nematodes or nematode suspensions using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 1 ml samples of whole blood using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 1-3 ml samples of amniotic fluid containing 0.5-2 x 106 cells using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 1, 10-15, or 20-30 Drosophila melanogaster using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 50 μl samples of dried human blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol is designed for purification of DNA from 50-75 mg dried or 100-200 mg fresh leaf tissue using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from a fecal swab or 1-2 g samples of fecal material using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit.
This protocol provides information about scaling of reagents required for purification of DNA from 0.5-700 mg solid tissue using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 0.1-0.3 mg samples of bone fragments using the Gentra Puregene Tissue Kit.
This protocol is designed for purification of DNA from 5 ml Gram-negative bacteria culture medium using the Gentra Puregene Cell Kit.
This protocol is designed for purification of DNA from 5 hair roots using the Gentra Puregene Tissue Kit.
This protocol provides information about scaling of reagents required for purification of DNA from 1-10 ml samples of fresh or frozen whole blood using the Gentra Puregene Blood Kit.
This protocol is designed for purification of DNA from 1 g samples of soil using the Gentra Puregene Cell 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
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

Do you have a protocol for the purification of archive-quality DNA from Eimeria oocysts using Gentra Puregene Kits?

Yes, please follow the Supplementary Protocol ' Purification of archive-quality DNA from Eimeria oocysts using Gentra Puregene Cell Kits ' (PG41).

 

 

FAQ ID -1952
What is the composition of RBC Lysis Solution?
The exact composition of RBC Lysis Solution is confidential. RBC Lysis Solution is a component of Gentra Puregene Blood Kits. RBC Lysis Solution selectively lyses human red blood cells leaving white blood cells intact. You can order RBC Lysis Solutin separately.
FAQ ID -2808
Do you have a protocol for the isolation of genomic DNA from bone?

Yes, we have the following protocols:

  • Isolation of genomic DNA from compact bone using the QIAamp DNA Mini Kit (QA02)
  • Isolation of genomic DNA from compact bone using the DNeasy Blood & Tissue Kit (DY01)
  • Purification of genomic DNA from bones using the QIAamp DNA Micro Kit (QA39)
  • Purification of archive-quality DNA from bone fragments using the Gentra Puregene Tissue Kit (PG38).
FAQ ID -908
Do you have a protocol for the purification of archive-quality DNA from buffy coat using Gentra Puregene Kits?
FAQ ID -1955
Do you have a protocol for the purification of archive-quality DNA from 1 ml of whole blood with the Gentra Puregene Tissue Kit?
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
Do you have a protocol for the purification of archive-quality DNA from fungus using Gentra Puregene Kits?
Do you have a protocol for the purification of archive-quality DNA from whole blood using Gentra Puregene Kits?
FAQ ID -1954
Do you have a protocol for the purification of archive-quality DNA from Gram-positive bacteria using Gentra Puregene Kits?
What is the composition of DNA Hydration Solution in Gentra Puregene Kits?

The composition of DNA Hydration Solution is as follows:

  • 10 mM Tris
  • 1 mM EDTA
  • pH 7–8

DNA Hydration Solution is included in QIAGEN’s Gentra Puregene Kits for DNA purification and can also be bought separately from QIAGEN.

FAQ ID -2813
Do you have a protocol for the purification of archive-quality DNA from Chlamydomonas using Gentra Puregene Kits?
What is the composition of Cell Suspension Solution?
The detailed composition of Cell Suspension Solution is confidential. Cell Suspension Solution is a component of the Gentra Puregene Yeast/Bacteria Kit. Essentially, it is an isotonic solution that prevents spheroblasts from bursting. Cell Suspension Solution can be ordered separately.
FAQ ID -2811
What is the composition of Protein Precipitation Solution?
The exact composition of Protein Precipitation Solution is confidential. Protein Precipitation Solution is a component of Gentra Puregene Kits for DNA purification. Essentially, this is a high-salt buffer that lowers the solubility of proteins. Protein Precipitation Solution can be ordered separately.
FAQ ID -2810
Do you have a protocol for the purification of DNA from fruit flies?

Using Gentra Puregene Cell kit:

 

• Purification of archive-quality DNA from up to 30 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG20)

• Purification of archive-quality DNA from 100 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG21)

• Purification of archive-quality DNA from 300–700 Drosophila melanogaster using the Gentra Puregene Cell Kit (PG22)

 

Using the QIAGEN Genomic tips:

• Isolation of genomic DNA from flies using the QIAGEN Genomic-tip 100/G (QG05)

FAQ ID -1970
Do you have a protocol for the purification of archive-quality DNA from blood smears using Gentra Puregene Kits?

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from blood smears using Gentra Puregene Cell Kits' (PG37).

 

 

FAQ ID -1948
Do you have a protocol for the purification of archive-quality DNA from 0.5–20 mg paraffin-embedded tissue using Gentra Puregene Kits?
Do you have a protocol for the purification of archive-quality DNA from fecal cells using Gentra Puregene Kits?
What is the composition and concentration of Glycogen Solution in Gentra Puregene Kits?
The exact composition of Glycogen Solution is confidential. The Glycogen concentration is 20mg/mL. Glycogen Solution is a component of Gentra Puregene Kits for DNA purification. During isopropanol precipitation, Glycogen Solution acts as a nucleic acid carrier and helps to efficiently precipitate small amounts of DNA. In addition, it facilitates visualization of the DNA pellet. Glycogen Solution can be purchased separately.

 

Note: Glycogen Solution does not work in combination with QIAGEN’s silica membrane technologies, e.g., RNeasy, DNeasy, QIAamp, etc. With these technologies, poly-A carrier RNA is used when dealing with low amounts of nucleic acids.

 

 

FAQ ID -2812
Do you have a protocol for the purification of archive-quality DNA from cerebrospinal fluid (CSF) using Gentra Puregene Kits?

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from 50 µl CSF using the Gentra Puregene Tissue Kit' (PG17).

 

 

 

 

 

FAQ ID -1967
Do you have a protocol for the purification of archive-quality DNA from hair roots using Gentra Puregene Kits?

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from hair roots using the Gentra Puregene Tissue Kit' (PG36).

 

 

 

FAQ ID -1947
Do you have a protocol for the isolation of genomic DNA from frozen clotted blood?

Yes, we have the following protocols:

  • Isolation of genomic DNA from frozen clotted whole blood using the QIAGEN Genomic-tip 100/G (QG02). TEST

You will need to prepare the required buffers according to the recipes in Appendix A of the QIAGEN Genomic DNA Handbook, or you can purchase the Genomic DNA Buffer Set containing pre-made solutions. Alternatively, the QIAGEN Blood & Cell Culture Midi Kit, containing Genomic-tips 100/G and buffers, can be used.

  • Purification of archive-quality DNA from clotted whole blood using Clotspin Baskets and the Gentra Puregene Blood Kit (PG03).
  • Purification of archive-quality DNA from clotted whole blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit (PG04).
  • Purification of DNA from clotted blood using the FlexiGene DNA Kit (FG01).
FAQ ID -900. Test
Do you have a protocol for the purification of archive-quality DNA from marine invertebrate tissue using Gentra Puregene Kits?
FAQ ID -1950
Do you have a protocol for the purification of archive-quality DNA from human dried blood using Gentra Puregene Kits?
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
Do you have a protocol for the purification of archive-quality DNA from umbilical cord blood using Gentra Puregene Kits?
Do you have a protocol for the purification of archive-quality DNA from nematodes using Gentra Puregene Kits?
FAQ ID -1951
What is the composition of Cell Lysis Solution?
The exact composition of Cell Lysis Solution is confidential. Cell Lysis Solution is a component of Gentra Puregene Kits for DNA purification. It functions by enabling cell lysis. Cell Lysis Solution can be purchased separately.
FAQ ID -2809
Do you have a protocol for rapid purification of archive-quality DNA from up to 9 x 10e6 cells using Gentra Puregene Kits?
What do you recommend for the cleanup of genomic DNA (gDNA)?

Using QIAGEN Genomic-tips

Protocol for DNA cleanup using Genomic-tips can be found in the 'Special Applications' section of the QIAGEN Genomic DNA Handbook.

 

Using QIAamp DNA Micro kit

Up to 10ug of gDNA can be cleaned using the cleanup protocol in the QIAamp DNA Micro Handbook.

 

Using Gentra Puregene Reagents

Gentra Puregene Handbook has protocols for removal of proteins and RNA from purified gDNA in Appendix C and Appendix D respectively.

FAQ ID -618
Do you have a protocol for the purification of archive-quality DNA from buccal brushes using Gentra Puregene Kits?

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from 5 buccal brushes using the Gentra Puregene Buccal Cell Kit' (PG15).

 

 

 

 

FAQ ID -1965
Do you have a protocol for the purification of archive-quality DNA from tissue fixed in ethanol or formalin using Gentra Puregene Kits?
Do I need to have EDTA in the buffer in which I am going to store my isolated genomic DNA?

EDTA chelates divalent cations which are required for nuclease activity. While the genomic DNA (gDNA) extracted using QIAGEN products, should not have any nuclease activity, it is possible to introduce nucleases during repeated long-term access of the DNA. EDTA helps to prevent any nuclease activity introduced after the genomic DNA extraction procedures.

However, if the gDNA is stored frozen at -20oC or -80oC, nuclease activity is much reduced. It may be possible to leave EDTA out of the storage buffer without negative consequences when samples are kept under these conditions, and when repeated freeze-thaw cycles are avoided.

We do recommend however that gDNA be stored in a neutral to a slightly basic buffered solution (e.g. 10 mM Tris-Cl pH 8.5 to 9.0) to prevent DNA degradation by acid hydrolysis. Note that deionized water mostly has an acidic pH.

FAQ ID -754
Do you have a protocol for the purification of archive-quality DNA from fish using Gentra Puregene Kits?
Do you have a protocol for the purification of archive-quality DNA from 100 to 5 x 10e8 cultured cells using Gentra Puregene Kits?