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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

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 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 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
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 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 90ml of buffered peptone water and homogenized for 1 minute in a 125µm filter stomacher bag. A 1.5ml 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
Tomatoe 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 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.

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Sample Type Promega External Comments
Bacteria – 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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Sample Type Promega External Comments
Bacteria – 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 #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

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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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Sample Type Promega External Comments
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 0.2µm 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 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 ~1ml aliquots and centrifuged at 14,000 × g for 5–7 minutes. The cell pellet was resuspended in 1 ml 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.

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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 (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.
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 15ml vials containing infected sodium-citrate whole blood at 3,000rpm for 10 minutes.
  2. Extract serum and leukocytes with 5ml pipettes.
  3. Dilute erythrocytes in 7 volumes of 0.15% saponin.
  4. Incubate at 37°C for 1 hour.
  5. Centrifuge at 3,000rpm for 40 minutes.
  6. Discard supernatant, taking care not to disturb the parasite pellet.
  7. 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 A260/280: 1.8.
(Personal communication)

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