Sample Types Processed with the Wizard(R) Genomic DNA Purification Kit

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DNA & RNA Purification

Sample Types Processed with the Wizard^® Genomic DNA Purification Kit

DNA from the following sample types has been successfully purified at Promega or by external laboratories using the Wizard^® Genomic DNA Purification Kit (Cat.# A1120, A1125, A1620).

Sample Types

Blood and blood cells

Mammalian tissues

Plant tissues

Cultured cells

Bacteria (Gram negative, Gram positive, other bacteria)

Yeast

Fungus

Other organisms and tissues

Many of these protocols have been communicated to Promega by external laboratories. Because these protocols have not been performed at Promega, they are intended to be guidelines that might require optimization.

This list will be updated as additional information becomes available. Contact Promega Technical Services (techserv@promega.com) for the latest information on available protocols.

Sample Types Processed Using the Wizard^® Genomic DNA Purification Kit.
Sample Type		Promega	External	Comments
Blood and blood cells
	Human whole blood	Yes	Yes	Protocols are provided in the Wizard^® Genomic DNA Purification Kit Technical Manual #TM050 for samples of up to 10 ml 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
	Mouse peripheral blood lymphocytes		Yes	Mehal, W.Z. and Crispe, I.N. (1998) J. Immunol. 161, 1686–93. PubMed ID #9712032
	Broiler (chicken) whole blood		Yes	Rossi, F. et al. (2005) Poult. Sci. 84, 1022–30. PubMed ID #16050119
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Sample Type		Promega	External	Comments
Mammalian tissues
	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 brain	Yes		Neural Notes (1995) Wizard^® Genomic DNA Purification Kit 1, 15.
	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.
	Meat (fermented sausage or minced meat)		Yes	Ten grams of fermented sausage or minced meat was diluted with 90 ml of buffered peptone water and homogenized for 1 minute in a 125 µm filter stomacher bag. A 1.5 ml aliquot of homogenate was centrifuged for 5 minutes at 14,000 × g at 4 °C, and genomic DNA was isolated. Martín, B. et al. (2006) Appl. Environ. Microbiol. 72, 6040–8. PubMed ID #16957227.
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Sample Type		Promega	External	Comments
Plant tissues
	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.
	Ancient moss		Yes	Marsic, D. et al. (1999) Extraction and amplification of DNA from an ancient moss. eNotes
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Sample Type		Promega	External	Comments
Cultured cells (Established cell lines are not listed.)		Yes	Yes	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: Zhang, S. et al. (2006) Cancer Res. 66, 8847–57. PubMed ID #16951202.
	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 300 µl 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.
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Sample Type		Promega	External	Comments
Bacteria
	Gram negative
	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	Genomic DNA was purified from colonies on chocolate agar. O’Neill, J.M. et al. (2003) J. Clin. Microbiol. 41, 3064–69. PubMed ID #12843045.
	Helicobacter pylori		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 1 ml 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.
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Sample Type		Promega	External	Comments
	Gram positive
	Staphylococcus epidermidis	Yes	Yes	Cells need to be treated with a lytic enzyme before lysis buffer addition. 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: Vandecasteele, S.J. et al. (2001) J. Bacteriol. 183, 7094–101. PubMed ID #11717267.
	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.
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	Other bacteria
	Treponema species		Yes	Cells were grown, centrifuged and washed three times with PBS. Protocol for Gram negative bacteria ("Isolating Genomic DNA from Gram Positive and Gram Negative Bacteria") was then followed. Additional Resource: Greene, S.R. and Stamm, L.V. (1998) Promega Notes 68, 30.
	Methanococcus jannaschii		Yes	Löwe, J. and Amos, L.A. (1998) Nature 391, 203–6. PubMed ID #16369025.
	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), 45 mM Tris (pH 8.0) and 9 mM 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 15 ml tube containing 9 ml of sterile PBS. After adding 4 mm 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 10 mg/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
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Sample Type		Promega	External	Comments
Yeast
	Saccharomyces cerevisiae	Yes	Yes	Yeast need to be treated with lyticase prior to adding lysis buffer. A protocol is provided in the Wizard^® Genomic DNA Purification Kit Technical Manual #TM050, "Isolating Genomic DNA from Yeast" section. Reference: Soteropoulos, P. and Perlin, D.S. (1998) J. Biol. Chem. 273, 26426–31. PubMed ID #9756876.
	Candida species, various		Yes	Yeast cells were incubated for 1 hour at 37 °C with 1 M sorbitol, 0.1 M 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.
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Fungus
	Madurella mycetomatis		Yes	Mycelia were sonicated and ground in liquid nitrogen before the addition of Nuclei Lysis Solution. Ahmed, A. et al. (2003) J. Clin. Microbiol. 41, 4537–41. PubMed ID #14532179.
	Aspergillus fumigatus		Yes	Cells were filtered through a 0.2 µm 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 50 mM Tris-HCl (pH 7.5), 50 mM 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 ~1 ml aliquots and centrifuged at 14,000 × g for 5–7 minutes. The cell pellet was resuspended in 1 ml of phosphate-buffered saline (0.01 M, pH 7.2) containing 1 mM 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.
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Sample Type		Promega	External	Comments
Other organisms and tissues
	Single flour beetles (Tribolium castaneum)		Yes	Lorenzen, M.D. et al. (2002) Genetics 160, 225–34. PubMed ID #11805058.
	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 (10 µm 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.
	Neocalanus species		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. 1. Centrifuge 15 ml vials containing infected sodium-citrate whole blood at 3,000 rpm for 10 minutes. 2. Extract serum and leukocytes with 5 ml pipettes. 3. Dilute erythrocytes in 7 volumes of 0.15% saponin. 4. Incubate at 37 °C for 1 hour. 5. Centrifuge at 3,000 rpm for 40 minutes. 6. Discard supernatant, taking care not to disturb the parasite pellet. 7. Resuspend the pellet in 15 ml of 1X PBS and centrifuge at 3,000 rpm 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 50 µl of DNA rehydration solution and incubated overnight at 4 °C. Yield: 2.5 µg per 3 × 10⁶ trophozoites. Purity A_260/280: 1.8. (Personal communication)
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