Puregene Kits

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

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Gentra Puregene Buccal Cell Kit (100)

Cat. No. / ID: 158845

For 100 preps: 100 Buccal Collection Brushes, RNase A Solution, Glycogen Solution, and Reagents

Kit

Gentra Puregene Kit

Puregene Kit

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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

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

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 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 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

Archive-quality DNA.

High-molecular-weight DNA from tissue.

Reliable RNA isolation from a single cell.

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.

See figures

Puregene DNA procedure.

Applications

DNA purified using 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 Puregene Kits
Features	Puregene Tissue Kit	Puregene Cell Kit	Puregene Blood Kit
Applications	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)	50–250 µl (varies)	250 µl – 1000 µl
Format	Scalable	Scalable	Scalable
Main sample type	Tissue samples, fixed and paraffin-embedded tissue	Cultured cells	Whole blood, buffy coat, body fluid
Processing	Manual	Manual	Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Genomic DNA	Genomic DNA	Genomic DNA
Sample amount	5–100 mg	1 x 10⁶ – 2.2 x 10⁷	300 µl – 10 ml / 1 x 10⁷ / 50 µl – 1000 µl
Technology	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 (+ DNA rehydration)	25–60 minutes (+ DNA rehydration)
Yield	Varies	7 µg / 1 million cells	35 µg / ml whole blood

Supporting data and figures

Exemplary size distribution using Puregene Cell Kit (Cells)

Exemplary size distribution of DNA from cells using the Puregene Cell Kit. Samples were analyzed with the Femto Pulse System and signals were normalized to maximum peak.

Resources

For purification of archive-quality DNA from human whole blood, bone marrow, buffy coat, body fluids, cultured cells, and tissue

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

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

FAQ ID -1952

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

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

Yes, please follow the Supplementary Protocol 'Scalable purification of archive-quality DNA from buffy coat using the Gentra Puregene Blood Kit' (PG02).

FAQ ID -1955

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from 1 ml whole blood with a body fluid protocol using the Gentra Puregene Tissue Kit' (PG42).

FAQ ID -1953

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from fungal tissue using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG23).

FAQ ID -1971

Yes, please follow the Supplementary Protocol 'Scalable purification of archive-quality DNA from whole blood using the Gentra Puregene Blood Kit' (PG01).

FAQ ID -1954

Yes, please choose from the Supplementary Protocols for various volumes of culture medium, or other starting materials, below:

Culture Medium:

Whole Blood:

'Purification of archive-quality DNA from Gram-positive bacteria in whole blood using the Gentra Puregene Yeast/Bacteria Kit' (PG32)

Soil:

'Purification of archive-quality DNA from Gram-positive soil bacteria using the Gentra Puregene Yeast/Bacteria Kit' (PG33)

FAQ ID -1946

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

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from Chlamydomonas using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG18).

FAQ ID -1968

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

DNA yield can differ between individual donors by up to 20-fold. Median human DNA yield, as determined by qPCR, from 290 different sample preparations with the Gentra Puregene Cell Kit was 16 µg DNA per ml saliva.

3822

Yes, please choose from the Supplementary Protocols for various volumes of culture medium, or other starting materials, below:

Culture medium:

Blood:

'Purification of archive-quality DNA from Gram-negative bacteria in whole blood using the Gentra Puregene Blood Kit' (PG30)

Soil:

'Purification of archive-quality DNA from Gram-negative soil bacteria using the Gentra Puregene Cell Kit' (PG31)

FAQ ID -1975

Protocols for automated DNA extraction from saliva stabilized with the PAXgene Saliva Collector are available for the QIAsymphony DNA Midi Kit with the QIAsymphony SP instrument or the QIAamp DNA Mini Kit with the QIAcube (Classic and Connect).
For manual extraction, supplementary protocols are available for the QIAamp DNA Mini and Gentra Puregene Cell Kits (see resources section).

3821

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

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

FAQ ID -1948

Yes, please follow the Supplementary Protocol 'Scalable purification of archive-quality DNA from 0.5–20 mg paraffin-embedded tissue using the Gentra Puregene Tissue Kit' (PG14).

FAQ ID -1964

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from fecal cells in suspension using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG24).

FAQ ID -1972

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

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

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

FAQ ID -1947

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

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from marine invertebrate tissue using the Gentra Puregene Tissue Kit' (PG39).

FAQ ID -1950

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from human dried blood using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG06).

FAQ ID -1959

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

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from umbilical cord blood using the Gentra Puregene Tissue Kit or Gentra Puregene Blood Kit' (PG05).

FAQ ID -1958

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from nematodes or nematode suspensions using the Gentra Puregene Tissue Kit' (PG40).

FAQ ID -1951

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

Yes, please follow the Supplementary Protocol 'Rapid purification of archive-quality DNA from 1–2 x 10e6, 3–5 x 10e6, or 6–9 x 10e6 cells using the Gentra Puregene Cell Kit' (PG19).

FAQ ID -1969

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

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

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from solid tissue fixed in ethanol or formalin using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG26).

FAQ ID -1974

Yes. A supplementary protocol for DNA extraction of 3 ml sample (2 ml saliva plus 1 ml stabilizing reagent) is available for the Gentra Puregene Cell Kit (for the supplementary protocol, see Resources section).

3826

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

Yes, please follow the Supplementary Protocol 'Purification of archive-quality DNA from 10–20 mg fish tissue using the Gentra Puregene Tissue Kit or Gentra Puregene Mouse Tail Kit' (PG25).

FAQ ID -1973

Yes, please follow the Supplementary Protocol 'Scalable purification of archive-quality DNA from 100 to 5 x 10e8 cultured cells using the Gentra Puregene Cell Kit' (PG09).

FAQ ID -1960