Wizard® Genomic DNA Purification Kit

Manual, Solution-Based DNA Extraction Method

Salting-out approach: 60-minute, scalable protocol
No chaotropic salts, silica membranes, or organic solvents are required, with minimal hazardous reagent handling
Preserves high-molecular-weight DNA integrity, which is important for applications that require intact, nonsheared DNA
Compatible with a wide variety of sample types and applications
Kit size based on number of isolations from whole blood; other sample types may vary

Catalog Number:

Size

100 isolations × 300μl 500 isolations × 300μl 100 isolations × 10ml

Catalog Number: A1120

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Catalog Number: A1125

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Catalog Number: A1620

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What Is the Salting-out Method for Genomic DNA Extraction?

Salting out is a solution-based DNA purification strategy in which high salt concentrations reduce protein solubility, causing them to precipitate out of solution while genomic DNA remains dissolved. The clarified DNA-containing supernatant is then recovered by alcohol precipitation. No silica columns or chaotropic reagents are required.

Salting out exploits the differential solubility of proteins and nucleic acids under high ionic strength conditions. At elevated salt concentrations—typically using concentrated sodium chloride (NaCl)—the ions compete with protein molecules for hydration by water, reducing the solvation shell around hydrophobic protein regions. This causes proteins to aggregate and precipitate as an insoluble pellet during centrifugation.

DNA, by contrast, remains soluble under these high-salt conditions because of its hydrophilic phosphate backbone. The result is a clear supernatant containing purified genomic DNA, free from the bulk of cellular proteins. The DNA is then concentrated and desalted by precipitation with isopropanol or ethanol, washed with 70% ethanol to remove residual salt, and resuspended in an aqueous rehydration buffer.

Why salting out preserves high-molecular-weight DNA

A key advantage of the salting-out approach over silica spin-column methods is the gentler handling of DNA. Column-based methods rely on vigorous mixing, centrifugation, and elution steps that can shear large DNA molecules. The Wizard® Genomic Kit’s solution-based protocol minimizes mechanical disruption, preserving fragments of high molecular weight—important for Southern blotting and restriction mapping where intact DNA is required.

The Wizard® Genomic DNA Purification Kit is based on the salting-out procedure originally described by Miller, Dykes and Polesky (1988) for extraction of DNA from human nucleated cells.

How Does the Wizard® Genomic DNA Purification Kit Work?

The Wizard® Genomic DNA Purification Kit includes four core reagents—Cell Lysis Solution, Nuclei Lysis Solution, Protein Precipitation Solution, and DNA Rehydration Solution—plus optional RNase A Solution. Together, they are used to execute the following workflow:

Cell lysis
RNase digestion (optional for blood; required for other sample types)
Protein precipitation
DNA precipitation and rehydration

RNase A treatment: optional or required by sample type

RNase A treatment removes RNA co-purified during cell lysis. Whether it is required depends on the sample type and downstream application.

Sample Type	RNase Treatment	Notes
Human whole blood (all volumes)	Optional	Not needed for most downstream applications
Mouse whole blood	Optional	—
Cell lines (K562, COS, NIH3T3, PC12, CHO)	Required	All cell line protocols require RNase step
Animal tissue (mouse liver, tail)	Required (liver); Optional (tail)	See Technical Manual #TM050
Insect cells (Sf9)	Required	—
Plant tissue (tomato leaf)	Required	—
Gram-negative and Gram-positive bacteria	Required	RNase added after 80°C lysis step
Yeast (S. cerevisiae)	Required	RNase added after DNA precipitation step

How Much Genomic DNA Does the Kit Yield?

DNA yield depends on sample type and the quantity of cells present. For human whole blood, typical yields are 5–15μg from 300μl, 25–50μg from 1ml, and 250–500μg from 10ml. Yields for other sample types vary by species and starting material; representative values are listed below.

The following yield data represent typical results using the standard protocols provided. Actual yields will vary depending on sample quality, storage conditions, and cell count.

Species/Material	Starting Amount	Typical DNA Yield	RNase Required
Human Whole Blood	300μl	5–15μg	Optional
	1.0ml	25–50μg	Optional
	10.0ml	250–500μg	Optional
Human Whole Blood (96-well)	50μl/well	0.2–0.7μg	Optional
Mouse Whole Blood (EDTA-treated)	300μl	6μg	Optional
Mouse Whole Blood (heparin-treated)	300μl	6–7μg	Optional
K562 (human cell line)	3 × 10⁶ cells	15–30μg	Required
COS (African green monkey)	1.5 × 10⁶ cells	10μg	Required
NIH3T3 (mouse)	2.25 × 10⁶ cells	9.5–12.5μg	Required
CHO (hamster)	1–2 × 10⁶ cells	6–7μg	Required
Mouse Liver	11mg	15–20μg	Required
Mouse Tail	0.5–1.0cm	10–30μg	Optional
Sf9 Insect Cells	5 × 10⁶ cells	16μg	Required
Tomato Leaf	40mg	7–12μg	Required
E. coli JM109	1ml overnight culture	20μg	Required
E. coli JM109	5ml overnight culture	75–100μg	Required
Staphylococcus epidermis	1ml overnight culture	6–13μg	Required
Saccharomyces cerevisiae	1ml overnight culture	4.5–6.5μg	Required

Overnight bacterial and yeast cultures are approximately 2 × 10⁹ cells/ml and 1.9 × 10⁸ cells/ml respectively.

How Does Solution-Based Extraction Compare to Silica Spin-Column DNA Purification?

Solution-based salting-out uses salt to selectively precipitate proteins while DNA stays in solution, avoiding silica membranes and chaotropic reagents. Silica spin-column methods adsorb DNA onto a membrane or matrix under chaotropic conditions and elute it in a small volume. The two approaches differ in DNA fragment size preservation, reagent requirements and workflow tradeoffs.

Feature	Solution-Based Salting Out (Wizard® Genomic)	Silica Spin-Column Methods
Mechanism	Salt precipitation of proteins; alcohol precipitation of DNA	Adsorption of DNA to silica membrane under chaotropic conditions
Chaotropic reagents	Not required	Required (guanidinium salts)
DNA fragment size	High-molecular-weight DNA preserved with gentle handling	Mechanical steps can introduce shearing
Elution volume	Flexible; resuspend in desired volume	Small, fixed elution volume; higher concentration
Protocol time	~60 minutes	~30–45 minutes (typically faster)
Hazardous reagents	Minimal; isopropanol and ethanol only	Guanidinium chloride or thiocyanate
Best for	Applications requiring HMW DNA (Southern blots, restriction mapping); broad sample variety	Routine PCR; concentrated DNA in small volumes

For most standard PCR and sequencing applications, either method produces DNA of sufficient quality. The salting-out approach has an advantage when DNA molecular weight integrity is critical or when minimizing chaotropic reagent handling is a priority.

For What Applications Is the Wizard® Genomic DNA Purification Kit Suitable?

The Wizard® Genomic DNA Purification Kit provides a simple, solution-based method for isolation of DNA from white blood cells, tissue culture cells, animal tissue, plant tissue, yeast and Gram-positive and Gram-negative bacteria. DNA purified with the Wizard® Genomic DNA Purification Kit is suitable for PCR amplification, restriction endonuclease digestion, Southern blotting, dot and slot blot hybridizations, and other standard molecular biology techniques requiring high-quality, intact genomic DNA.

Learn more about our custom options for this product at: www.promega.com/custom/

Sample types processed with the Wizard® Genomic DNA Purification Kit

The following tables document validated sample types for the Wizard® Genomic DNA Purification Kit, with references to protocols in Technical Manual #TM050 and peer-reviewed publications demonstrating use with external samples. This is one of the most extensively validated genomic DNA extraction kits across species and sample types in the published literature.

Blood

Sample Type	Promega	External	Comments
Human whole blood	Yes	Yes	Protocols are provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050 for samples of up to 10ml blood or in 96-well plates (“Isolating Genomic DNA from Whole Blood” sections). The protocol works with the following anticoagulants: EDTA, heparin and citrate. Reference: Rizzo, W.B., Carney, G. and Lin, Z. (1999) Am. J. Hum. Genet. 65, 1547–60. PubMed ID 10577908
Mouse whole blood	Yes	—	A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050.
Human peripheral blood mononuclear cells	—	Yes	Tan, A. et al. (2006) Clin. Chem. 52, 2250–7. PubMed ID 17040960
Bovine whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Horse whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Sheep whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Antelope whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Dog whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Cat whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348
Rabbit whole blood	—	Yes	Walker, J.A. et al. (2003) Anal. Biochem. 316, 259–69. PubMed ID 12711348

Mammalian Tissues

Sample Type	Promega	External	Comments
Mouse liver	Yes	Yes	Tissue needs to be homogenized prior to isolation. A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, “Isolating Genomic DNA from Tissue Culture Cells and Animal Tissue” section. Reference: Barazzoni, R. et al. (2005) Endocrinology 146, 2098–106. PubMed ID 15618355.
Mouse tail	Yes	Yes	Tail needs to be treated with proteinase K before purification. A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, “Isolating Genomic DNA from Tissue Culture Cells and Animal Tissue” section. Reference: Nielsen, J.V. et al. (2007) Development 134, 1133–40. PubMed ID 17301088.
Mouse brain	—	Yes	Sinz, E.H. et al. (1999) J. Clin. Invest. 104, 647–56. PubMed ID 10487779.
Mouse lymphoma	—	Yes	Yu, Z. et al. (2006) Cancer Res. 66, 755–62. PubMed ID 16424006.
Rat skeletal muscle, liver and heart	—	Yes	Barazzoni, R. et al. (2000) J. Biol. Chem. 275, 3343–7. PubMed ID 10652323.
Human bone marrow aspirate	—	Yes	Yu, Z. et al. (2006) Cancer Res. 66, 755–62. PubMed ID 16424006.
Human liver and testicle	—	Yes	Maduro, M.R. et al. (2003) Mol. Hum. Reprod. 9, 61–8. PubMed ID 12569174.
Human colon	—	Yes	Tost, J. et al. (2003) Nucleic Acids Res. 31, e50. PubMed ID 12711695.

Plant Tissues

Sample Type	Promega	External	Comments
Tomato leaf	Yes	—	Leaf needs to be frozen in liquid nitrogen and ground with a mortar and pestle prior to adding lysis buffer. A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, “Isolating Genomic DNA from Plant Tissue” section.
Arabidopsis	—	Yes	Vorwerk, S. (2001) Wizard® Genomic DNA Purification Kit and the isolation of plant genomic DNA. eNotes
Maize	—	Yes	Vorwerk, S. (2001) Wizard® Genomic DNA Purification Kit and the isolation of plant genomic DNA. eNotes
Lemna minor (aquatic plant)	—	Yes	Stout, L.M. and Nüsslein, K. (2005) Appl. Environ. Microbiol. 71, 2484–92. PubMed ID 15870338.
Medicago truncatula	—	Yes	Total DNA was isolated from roots, which may include various mycorrhizal fungi. Kosuta, S. et al. (2003) Plant Physiol. 131, 952–62. PubMed ID 12644648.
Tetrahymena thermophila	—	Yes	Followed plant DNA protocol (“Isolating Genomic DNA from Plant Tissue”) but skipped freezing in liquid nitrogen and grinding with a mortar and pestle. Jacobs, M.E. et al. (2006) Eukaryot. Cell 5, 1990–2000. PubMed ID 17012537.

Cultured Cells

Sample Type	Promega	External	Comments
Drosophila D.Mel-2 embryonic cells	—	Yes	Roberti, M. et al. (2006) Nucleic Acids Res. 34, 2109–16. PubMed ID 16648357.
Mouse embryonic stem cells	—	Yes	Oka, M. et al. (2006) J. Biol. Chem. 281, 9901–8. PubMed ID 16439359.
Monkey primary fibroblasts, virus-infected	—	Yes	Cells were lysed using 300μl of Nuclei Lysis Solution with proteinase K and incubating overnight at 37°C. Mansfield, K.G. et al. (1999) J. Virol. 73, 10320–8. PubMed ID 10559350.
Vero cells infected with HSV-1	—	Yes	Both viral and genomic DNA were isolated. Bower, J.R. et al. (1999) J. Virol. 73, 3843–53. PubMed ID 10196279.
CD45RA+ T cells from cord blood	—	Yes	Samples of 2–4 × 10⁶ cells were pelleted, washed with PBS, snap frozen in liquid nitrogen and frozen at −70°C prior to using the Wizard® Genomic DNA Purification Kit. Soares et al. (1998) J. Immunol. 161, 5909–17. PubMed ID 9834071.

Bacteria: Gram-Negative

Sample Type	Promega	External	Comments
Enterobacter cloacae	Yes	—	A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, “Isolating Genomic DNA from Gram Positive and Gram Negative Bacteria” section.
Escherichia coli	Yes	Yes	A protocol is provided in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, “Isolating Genomic DNA from Gram Positive and Gram Negative Bacteria” section. Reference: Zheng, W. et al. (2001) EMBO J. 20, 1164–72. PubMed ID 11230139.
Nitrobacter winogradskyi	—	Yes	Starkenburg, S.R. et al. (2006) Appl. Environ. Microbiol. 72, 2050–63. PubMed ID 16517654.
Yersinia pestis	—	Yes	Flashner, Y. et al. (2004) Infect. Immun. 72, 908–15. PubMed ID 14742535.
Burkholderia species	—	Yes	Moore, R.A. et al. (2004) Infect. Immun. 72, 4172–87. PubMed ID 15213162.
Salmonella enterica	—	Yes	Nair, S. et al. (2004) J. Bacteriol. 186, 3214–23. PubMed ID 15126484.
Field bacteria samples, non-Agrobacterium	—	Yes	Weller, S.A. et al. (2004) Appl. Environ. Microbiol. 70, 2779–85. PubMed ID 15128532.
Haemophilus influenzae	—	Yes	Ottemann, K.M. and Lowenthal, A.C. (2002) Infect. Immun. 70, 1984–90. PubMed ID 11895962.
Bordetella bronchiseptica	—	Yes	Brockmeier, S.L. et al. (2002) Infect. Immun. 70, 481–90. PubMed ID 11796573.
Leptospira kirschneri	—	Yes	Matsunaga, J. et al. (2002) Infect. Immun. 70, 323–34. PubMed ID 11748198.
Klebsiella pneumoniae	—	Yes	Yigit, H. et al. (2001) Antimicrob. Agents Chemother. 45, 1151–61. PubMed ID 11257029.
Marine Geobacteraceae	—	Yes	Bacteria colonizing energy-harvesting anodes were scraped off, organically extracted and purified using the Wizard® Genomic DNA Purification Kit. Bond, D.R. et al. (2002) Science 295, 483–5. PubMed ID 11799240.
Serratia marcescens	—	Yes	Peccia, J. and Hernandez, M. (2001) Appl. Environ. Microbiol. 67, 4225–32. PubMed ID 11526027.
Borrelia burgdorferi	—	Yes	Chaconas, G. et al. (2001) EMBO J. 20, 3229–37. PubMed ID 11406599.
Campylobacter jejuni	—	Yes	Dorrell, N. et al. (2001) Genome Res. 11, 1706–15. PubMed ID 11591647.
Desulfovibrio desulfuricans	—	Yes	Rapp-Giles, B.J. et al. (2000) Appl. Environ. Microbiol. 66, 671–7. PubMed ID 10653734.
Vibrio cholerae	—	Yes	Stine, O.C. et al. (2000) Infect. Immun. 68, 7180–5. PubMed ID 11083852.
Vibrio harveyi	—	Yes	Zhou, W. et al. (1999) Biochemistry 38, 16246–52. PubMed ID 10587447.
Paracoccus denitrificans	—	Yes	Shearer, N. et al. (1999) J. Bacteriol. 181, 6907–13. PubMed ID 10559155.
Campylobacter species	—	Yes	Before genomic DNA isolation, the bacteria were washed in TE buffer and resuspended in 1ml of TE buffer at ~1 × 10⁹ cells/ml. Duim, B. et al. (1999) Appl. Environ. Microbiol. 65, 2369–75. PubMed ID 10347015.
Sphingomonas paucimobilis	—	Yes	Jiang, S.C. et al. (1998) Appl. Environ. Microbiol. 64, 535–42. PubMed ID 9464390.
Flavobacterium species	—	Yes	Jiang, S.C. et al. (1998) Appl. Environ. Microbiol. 64, 535–42. PubMed ID 9464390.

Bacteria: Gram-Positive

Sample Type	Promega	External	Comments
Bacillus anthracis	—	Yes	Pomerantsev, A.P. et al. (2006) Infect. Immun. 74, 682–93. PubMed ID 16369025.
Bacillus cereus	—	Yes	van Schaik, W. et al. (2004) J. Bacteriol. 186, 316–25. PubMed ID 14702299.
Bacillus subtilis	—	Yes	Ben-Yehuda, S. and Losick, R. (2002) Cell 109, 257–66. PubMed ID 12007411.
Staphylococcus, 12 strains	—	Yes	Li, Y. et al. (2003) J. Clin. Microbiol. 41, 3481–6. PubMed ID 12904342.
Lactobacillus acidophilus	—	Yes	Li, Y. et al. (2003) J. Clin. Microbiol. 41, 3481–6. PubMed ID 12904342.
Actinomyces naeslundii	—	Yes	Li, Y. et al. (2003) J. Clin. Microbiol. 41, 3481–6. PubMed ID 12904342.
Streptococcus pneumoniae	—	Yes	Brown, J.S. et al. (2002) Infect. Immun. 70, 4389–98. PubMed ID 12117949.

Bacteria: Other

Sample Type	Promega	External	Comments
Methanococcus jannaschii	—	Yes	Löwe, J. and Amos, L.A. (1998) Nature 391, 203–6. PubMed ID 9428770.
Methanosarcina mazei	—	Yes	Lange, M. et al. (2000) Appl. Environ. Microbiol. 66, 1796–1800. PubMed ID 10788341.
Mycoplasma hyorhinis	—	Yes	Cells were collected and lysed in 1% sodium dodecyl sulfate (SDS), 45mM Tris (pH 8.0) and 9mM EDTA for 10 minutes at 50°C, then incubated for 10 minutes at 37°C. The Wizard® Genomic DNA Purification Kit was subsequently used for genomic DNA extraction. Citti, C. et al. (2000) J. Bacteriol. 182, 1356–63. PubMed ID 10671459.
Mycobacterium parafortuitum	—	Yes	Cells were treated with proteinase K and lysozyme for 1 hour, centrifuged and resuspended in Nuclei Lysis Solution. Glass beads were added, the glass bead-cell lysate shaken for 5 minutes, incubated at 65°C for 1 hour and the glass bead-lysate shaken again for 3 minutes. The protocol then followed the bacterial method of the Wizard® Genomic DNA Purification Kit (“Isolating Genomic DNA from Gram Positive and Gram Negative Bacteria”). Peccia, J. and Hernandez, M. (2001) Appl. Environ. Microbiol. 67, 4225–32. PubMed ID 11526027.
Intestinal microflora	—	Yes	Chicken intestines were removed and the contents of each segment (duodenum, jejunum, ileum, and cecum) were inverted into a sterile 15ml tube containing 9ml of sterile PBS. After adding 4mm glass beads, the samples were vortexed for 3 minutes, and the debris was removed by centrifuging at 700 × g for 1 minute. The supernatant was collected and centrifuged at 12,000 × g for 5 minutes. The pellet was washed twice with PBS and stored at −20°C until DNA extraction. This frozen pellet was resuspended in EDTA and treated with lysozyme (at a final concentration of 10mg/ml) for 45 minutes at 37°C. The bacterial genomic DNA was isolated with the Wizard® Genomic DNA Purification Kit. Amit-Romach, E., Sklan, D. and Uni, Z. (2004) Poult. Sci. 83, 1093–98. PubMed ID 15285498.

Yeast

Sample Type	Promega	External	Comments
Candida species, various	—	Yes	Yeast cells were incubated for 1 hour at 37°C with 1M sorbitol, 0.1M EDTA, 0.1% (w/v) zymolyase-100T (Seikaguku Corp.), and 1% (v/v) 2-mercaptoethanol adjusted to pH 7.5. Chromosomal DNA was then isolated using the Wizard® Genomic DNA Purification Kit. Park, S. et al. (2000) J. Clin. Microbiol. 38, 2829–36. PubMed ID 10921935.

Fungus

Sample Type	Promega	External	Comments
Aspergillus fumigatus	—	Yes	Cells were filtered through a 0.2μm SFCA filter, frozen on dry ice, then ground with a mortar and pestle. The ground cells were incubated in Nuclei Lysis Solution at 37°C for 1 hour before purifying with the Wizard® Genomic DNA Purification Kit, “Isolating Genomic DNA from Tissue Culture Cells and Animal Tissue” section. Burghoorn, H.P. et al. (2002) Antimicrob. Agents Chemother. 46, 615–24. PubMed ID 11850239.
Nectria haematococca	—	Yes	Mycelia were ground in liquid nitrogen and lyophilized before lysing in a modified lysis buffer consisting of 50mM Tris-HCl (pH 7.5), 50mM EDTA (pH 8.0), 3% SDS and 1% mercaptoethanol. Wu, Q. et al. (1998) Mol. Biol. Cell 9, 89–101. PubMed ID 9436993.
Pneumocystis carinii	—	Yes	Sputum samples were dispensed into ~1ml aliquots and centrifuged at 14,000 × g for 5–7 minutes. The cell pellet was resuspended in 1ml of phosphate-buffered saline (0.01M, pH 7.2) containing 1mM EDTA (PBS-EDTA), washed twice in PBS-EDTA, centrifuged and stored at −80°C for later DNA extraction. DNA was prepared using the Wizard® Genomic DNA Purification Kit, “Isolating Genomic DNA from Whole Blood” section. Beard, C.B. et al. (2000) Emerg. Infect. Dis. 6, 265–72. PubMed ID 10827116.

Other Organisms and Tissues

Sample Type	Promega	External	Comments
Drosophila, adult fly	—	Yes	To prepare genomic DNA from adult D. virilis and D. melanogaster, 100 adult flies were homogenized and the DNA purified using the Wizard® Genomic DNA Purification Kit. Hanrahan, C.J. et al. (2000) Genetics 155, 1149–60. PubMed ID 10880477.
Frog skin	—	Yes	Skin biopsies from Amphibiocystidium ranae (10μm sections) were deparaffinized twice in xylene and centrifuged at high speed. The tissue pellet was washed with 95% and 70% ethanol, then dried, and the genomic DNA was extracted using the Wizard® Genomic DNA Purification Kit. Pereira, C.N. et al. (2005) J. Clin. Microbiol. 43, 192–8. PubMed ID 15634971.
Cichlid fin and whole fish	—	Yes	Genomic DNA was isolated from 1–2 mm² ethanol-preserved Neolamprologus pulcher finclip samples or whole offspring using the Wizard® Genomic DNA Isolation Kit, “Isolating Genomic DNA from Tissue Culture Cells and Animal Tissue” section, with small modifications (not listed). Heg, D. et al. (2006) Behav. Ecol. 17, 419–29.
Plasmodium vivax	—	Yes	Researchers used a modified animal tissue/tail snip protocol. Sawabe, T. and Ikeda, T. (2005) Wizard® Genomic DNA Purification Kit provides high-quality genomic DNA template for molecular phylogenetic studies on Copepod crustaceans. eNotes
Plasmodium vivax	—	Yes	To isolate the parasite from whole blood, a modified protocol was used. Centrifuge 15ml vials containing infected sodium-citrate whole blood at 3,000rpm for 10 minutes. Extract serum and leukocytes with 5ml pipettes. Dilute erythrocytes in 7 volumes of 0.15% saponin. Incubate at 37°C for 1 hour. Centrifuge at 3,000rpm for 40 minutes. Discard supernatant, taking care not to disturb the parasite pellet. Resuspend the pellet in 15ml of 1X PBS and centrifuge at 3,000rpm for 10 minutes. Discard the supernatant, and repeat the procedure twice. Start with the Nuclei Lysis Solution addition step in the “Isolating Genomic DNA from Whole Blood” section of the Wizard® Genomic DNA Purification Kit Technical Manual #TM050. (Personal communication)
Amoeba	—	Yes	Trophozoites of clone A of Entamoeba histolytica (strain HM1:IMSS) were “axenically cultured” in TYI-S33 medium. Trophozoites (3 × 10⁶) were processed as described in the Wizard® Genomic DNA Purification Kit Technical Manual #TM050, following the protocol for isolation of genomic DNA from tissue culture cells (“Isolating Genomic DNA from Tissue Culture Cells and Animal Tissue” section). DNA pellet was dissolved with 50μl of DNA rehydration solution and incubated overnight at 4°C. Yield: 2.5μg per 3 × 10⁶ trophozoites. Purity A_260/280: 1.8. (Personal communication)

Protocols

Complete Protocol

Wizard® Genomic DNA Purification Kit Technical ManualPDF (1019 KB) – English

Quick Protocols

Wizard Genomic DNA Purification Kit FB022PDF (432 KB) – English

Frequently Asked Questions

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What is the principle of salting-out DNA purification?

Salting out exploits the difference in solubility between proteins and DNA at high ionic strength. When concentrated NaCl (the Protein Precipitation Solution) is added to a cell lysate, it reduces water availability for protein hydration, causing proteins to aggregate and precipitate. DNA remains in solution due to its hydrophilic phosphate backbone. The proteins are pelleted by centrifugation, and DNA is recovered from the supernatant by isopropanol precipitation. This method was originally described by Miller, Dykes and Polesky (1988) Nucl. Acids Res. 16: 1215.

Why are red blood cells lysed first in whole-blood DNA extraction?

Red blood cells are anucleate—they contain no nucleus and therefore no genomic DNA. Lysing them selectively with Cell Lysis Solution removes the majority of cell volume before WBC lysis, concentrating the white blood cells that are the actual source of nuclear DNA. This makes the subsequent lysis and protein precipitation steps more efficient. The WBC pellet must be fully vortexed after RBC removal to ensure complete resuspension—incomplete resuspension is the most common cause of poor yield in blood protocols.

How does high salt precipitate proteins during salting-out extraction?

At high ionic strength, Na⁺ and Cl⁻ ions from the Protein Precipitation Solution compete with protein molecules for hydration by surrounding water. This strips the hydration shell from protein surfaces, exposing hydrophobic regions that aggregate with other protein molecules. The resulting aggregates are insoluble and form a dark brown pellet during centrifugation. DNA, which has a strongly hydrophilic phosphate backbone, remains in solution under these same conditions.

What is the role of Cell Lysis Solution and Nuclei Lysis Solution?

Cell Lysis Solution is used exclusively in blood protocols to selectively lyse red blood cells, which are more osmotically fragile than white blood cells. It does not lyse white blood cells or their nuclei. Nuclei Lysis Solution is used in all protocols to disrupt white blood cell and nuclear membranes (or cell membranes in non-blood samples), releasing genomic DNA into solution. The solution should become viscous after addition, indicating DNA release. If cell clumps remain after mixing, incubation at 37°C is recommended until disrupted.

Why does the DNA pellet need to be washed with 70% ethanol?

The DNA pellet after isopropanol precipitation retains residual salts from the Protein Precipitation Solution. These salts can inhibit downstream enzymes including polymerases and restriction endonucleases. A wash with 70% ethanol removes salt without dissolving the DNA pellet. Complete removal of ethanol before resuspension is important—residual ethanol can inhibit PCR. The DNA pellet is very loose at the wash step; use extreme care when aspirating to avoid pellet loss.

Why might no protein pellet be visible after centrifugation?

Two causes are identified in Technical Manual #TM050, Section 4: (1) The sample was not cooled to room temperature before adding Protein Precipitation Solution. The sample must be at room temperature for at least 5 minutes, or chilled on ice for 5 minutes, before proceeding. (2) The Protein Precipitation Solution was not thoroughly mixed with the nuclear lysate. Always vortex vigorously for 10–20 seconds to ensure complete mixing. If no pellet appears after these steps, centrifuge again for 3 minutes at 13,000–16,000 × g.

What causes a DNA pellet that is difficult to dissolve?

According to Technical Manual #TM050, Section 4, a pellet that is difficult to dissolve is most commonly caused by overdrying. Air-dry the pellet for only 10–15 minutes—do not leave it to dry for extended periods. If the pellet is already overdried, rehydrate by incubating at 65°C for 1 hour with periodic gentle mixing, then leave the sample at room temperature or 4°C overnight. Do not leave the DNA at 65°C overnight. Incomplete mixing during the rehydration step can also produce a pellet that appears undissolved; mix periodically throughout the rehydration incubation.

Can frozen blood samples be used with the Wizard® Genomic Kit?

Yes, but yields may be lower than from fresh blood. When using frozen blood, repeat the Cell Lysis Solution incubation and centrifugation steps until the pellet appears white, as additional Cell Lysis Solution may be needed to fully remove lysed RBCs. For optimal results, use fresh blood or samples stored at 2–8°C for no more than 5 days. DNA from blood samples stored for more than 5 days at 2–8°C may show reduced yield.

What should I do if blood is clotted?

Clotted blood cannot be processed with this kit. Clotting indicates the sample was stored improperly, the blood was not thoroughly mixed with anticoagulant at collection, or the incorrect tube type was used. Discard the clotted sample and draw a new blood sample using EDTA-, heparin-, or citrate-treated anticoagulant tubes. Blood must be collected in anticoagulant tubes; clot-activator (gold-top SST) tubes are not compatible with this protocol.

How is DNA concentration and purity assessed after Wizard® Genomic extraction?

DNA concentration and purity are typically assessed by spectrophotometry. An A₂₆₀/A₂₈₀ ratio between 1.7 and 1.9 indicates acceptably pure DNA; values below 1.7 suggest protein contamination, and values above 1.9 may indicate RNA contamination (if RNase step was skipped). An A₂₆₀/A₂₃₀ ratio above 1.8 indicates low salt or carbohydrate contamination. For precise quantification of low-concentration samples, fluorometric methods (Qubit®, PicoGreen®) are more accurate than UV absorbance.

How much genomic DNA can be expected from a 300μl whole-blood sample?

The typical yield from 300μl of human whole blood is 5–15μg (Technical Manual #TM050). Yield depends primarily on the white blood cell count of the donor sample. Human adult blood contains approximately 4,000–11,000 WBCs per microliter; lymphopenic samples will produce lower yields. Resuspension in 100μl DNA Rehydration Solution gives a typical working concentration of approximately 50–150ng/μl. Yields from mouse blood (300μl) are typically ~6μg for both EDTA- and heparin-collected samples.

What anticoagulants are compatible with the Wizard® Genomic Kit blood protocol?

The blood protocol is compatible with EDTA, heparin, and sodium citrate anticoagulants. EDTA (purple-top tubes) is the standard for DNA extraction and is preferred for most molecular biology applications. Heparin-collected samples are compatible but heparin is a known inhibitor of Taq polymerase and PCR (Beutler et al., 1990 BioTechniques 9: 166); if PCR is the intended downstream application, EDTA-collected blood is strongly preferred. Anticoagulant blood samples may be stored at 2–8°C for up to two months, but DNA yield will decrease with storage time.

Can the Wizard® Genomic Kit be used for bacteria and yeast DNA extraction?

Yes. Gram-negative bacteria (e.g., E. coli) require only the standard Nuclei Lysis Solution step, using an 80°C incubation for 5 minutes to lyse cells. Gram-positive bacteria require pretreatment with lysozyme (or lysozyme + lysostaphin for Staphylococcus) to weaken the thick peptidoglycan cell wall before lysis. Yeast require pretreatment with lyticase to digest the glucan cell wall. After the pretreatment step, protein precipitation and DNA recovery proceed identically to the mammalian cell protocol. RNase treatment is required for all bacterial and yeast samples.

Specifications

Catalog Number:

What's in the box?

Item	Part #	Size	Available Separately
Cell Lysis Solution	A793A	1 × 100ml	View Product
Nuclei Lysis Solution	A7941	1 × 50ml	View Product
Protein Precipitation Solution	A795A	1 × 25ml	View Product
DNA Rehydration Solution	A796A	1 × 50ml	View Product
RNase A Solution	A797A	1 × 250μl	View Product

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

What's in the box?

Item	Part #	Size	Available Separately
Cell Lysis Solution	A793A	5 × 100ml	View Product
Nuclei Lysis Solution	A7941	5 × 50ml	View Product
Protein Precipitation Solution	A795A	5 × 25ml	View Product
DNA Rehydration Solution	A796A	2 × 50ml	View Product
RNase A Solution	A797A	5 × 250μl	View Product

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

What's in the box?

Item	Part #	Size
Cell Lysis Solution (Genomic Purification)	A7933	3 × 1 liter
Nuclei Lysis Solution	A7943	1 × 1 liter
Protein Precipitation Solution	A7953	1 × 350ml
DNA Rehydration Solution	A7963	3 × 50ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Resources

Articles

Distribution of honey bee mitochondrial DNA haplotypes in an Italian region where a legislative act is protecting the Apis mellifera ligustica subspecies

Other Resources

DNA Purification

Product Citations

Recent publications that mention the use of this product.

View PubMed Citations

Wizard® Genomic DNA Purification Kit

Manual, Solution-Based DNA Extraction Method

What Is the Salting-out Method for Genomic DNA Extraction?

Why salting out preserves high-molecular-weight DNA

How Does the Wizard® Genomic DNA Purification Kit Work?

RNase A treatment: optional or required by sample type

How Much Genomic DNA Does the Kit Yield?

How Does Solution-Based Extraction Compare to Silica Spin-Column DNA Purification?

For What Applications Is the Wizard® Genomic DNA Purification Kit Suitable?

Sample types processed with the Wizard® Genomic DNA Purification Kit

Protocols

Complete Protocol

Quick Protocols

Frequently Asked Questions

What is the principle of salting-out DNA purification?

Why are red blood cells lysed first in whole-blood DNA extraction?

How does high salt precipitate proteins during salting-out extraction?

What is the role of Cell Lysis Solution and Nuclei Lysis Solution?

Why does the DNA pellet need to be washed with 70% ethanol?

Why might no protein pellet be visible after centrifugation?

What causes a DNA pellet that is difficult to dissolve?

Can frozen blood samples be used with the Wizard® Genomic Kit?

What should I do if blood is clotted?

How is DNA concentration and purity assessed after Wizard® Genomic extraction?

How much genomic DNA can be expected from a 300μl whole-blood sample?

What anticoagulants are compatible with the Wizard® Genomic Kit blood protocol?

Can the Wizard® Genomic Kit be used for bacteria and yeast DNA extraction?

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Resources

Articles

Other Resources

Related Products

Similar Products

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ReliaPrep™ Blood gDNA Miniprep System

ReliaPrep™ gDNA Tissue Miniprep System

SV Total RNA Isolation System

Frequently Used With

Molecular Biology Lab Guide

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GoTaq® G2 Master Mixes

QuantiFluor® ONE dsDNA System

Quantus™ Fluorometer and NGS Starter Package

Nuclei Lysis Solution

Product Citations

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