Wizard® Genomic DNA Purification Kit

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	For blood DNA isolation, blood must be collected in EDTA, heparin or citrate anticoagulant tubes to prevent clotting.

Protocol tips
For blood DNA isolation, blood must be collected in EDTA, heparin or citrate anticoagulant tubes to prevent clotting.

Publication protocol

DNA and cell lines
Genomic mouse DNA samples obtained from Jackson Laboratories (Bar Harbor, ME, USA) include 37 inbred mice DNA samples, 1 recombinant inbred mouse sample, and 10 wild-derived mice DNA samples. DNA from 15 wild-caught mice (courtesy of Dr. Michael Nachman from the University of Arizona, USA) collected in Tucson, AZ was used for heterozygosity studies. Genomic DNA from mouse (male and female CD1/ICR), hamster (Syrian golden hamster, Chinese hamster), rat (Fischer, Wistar, Sprague–Dawley), gerbil, pig, baboon, rhesus, and cynomolgus monkey were obtained from Zyagen (San Diego, CA, USA). TN1 cells used in stability studies (courtesy of Dr. Anne Plant from NIST) are a stable transfected NIH3T3 cell line expressing green fluorescent protein (GFP) (parent NIH3T3 cells obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) in 2003). Cell lines, derived from mouse, Chinese hamster, human, and African green monkey, were obtained from ATCC and their respective growth requirements are described in Table . All cells were grown in a humidified 5 % CO balanced-air atmosphere at 37 °C. For DNA extraction purposes, all cell lines were harvested at passage 3. Adherent cell lines were harvested using 0.25 % trypsin/0.53 mM EDTA solution (ATCC). Trypsin activity was quenched by the addition of an equivalent volume of growth medium (0.1 % soybean inhibitor (Invitrogen) used to neutralize trypsin for MCF 10A cells) and one million cells from each cell line were counted using the Multisizer 3 Coulter Counter (Beckman Coulter, Brea, CA, USA). RAW 264.7 cells were harvested using a cell scraper. The Wizard DNA Extraction kit (Promega, Madison, WI, USA) was used to isolate DNA from harvested cells. DNA was quantified using the Synergy Mx plate reader and Take3 plate (BioTek, Winooski, VT, USA) at an absorbance of 260 nm. To study STR marker stability over increasing passage number, duplicate 25 cm tissue culture flasks of L929 cells were carried independently, and one million cells were harvested at passage numbers 2, 4, 9, 14, 19, 22, 26, 29, 31, 37, 41, and 44. Duplicate 25 cm flasks were also carried for NIH3T3 cells which were carried independently, and one million cells were harvested from passage numbers 5, 7, 10, 15, 20, 23, 26, 32, 35, 40, 43, and 45.

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Manufacturer protocol

Download the product protocol from Promega for Wizard® Genomic DNA Purification Kit below.

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