Wizard Genomic DNA Purification Kit Protocol - Department of ...

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Large-Scale Isolation (maxiprep) Product Size Cat.# Wizard ® Genomic DNA Purification Kit 100 isolations A1620 Each system contains sufficient reagents for 100 isolations of genomic DNA from 10ml of whole blood samples. Includes: • 3L Cell Lysis Solution • 1L Nuclei Lysis Solution • 350ml Protein Precipitation Solution • 150ml DNA Rehydration Solution Note: Cat.# A1620 does not include RNase Solution. Items Available Separately Product Size Cat.# Cell Lysis Solution 1L A7933 Nuclei Lysis Solution 1L A7943 Protein Precipitation Solution 350ml A7953 DNA Rehydration Solution 50ml A7963 RNase A (4mg/ml) 1ml A7973 Storage Conditions: Store the Wizard ® Genomic DNA Purification Kit at room temperature (22–25°C). See product label for expiration date. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.com Printed in USA. Part# TM050 Revised 12/10 Page 3
Table 1. DNA Yields from Various Starting Materials. Species and Material Human Whole Blood (Yield depends on the quantity of white blood cells present) 96-well plate (Process as little as 20µl/well; see Table 2.) Mouse Whole Blood EDTA (4%) treated Heparin (4%) treated 96-well plate Cell Lines K562 (human) COS (African green monkey) NIH3T3 (mouse) PC12 (rat pheochromocytoma) CHO (hamster) Animal Tissue Mouse Liver Mouse Tail Amount of Starting Material 300µl 1.0ml 10.0ml 50µl/well 300µl 300µl 50µl/well 3 × 10 6 cells 1.5 × 10 6 cells 2.25 × 10 6 cells 8.25 × 10 6 cells 1–2 × 10 6 cells 11mg 0.5–1.0cm of tail Typical DNA Yield 5–15µg 25–50µg 250–500µg 0.2–0.7µg 6µg 6–7µg 0.2–0.7µg 15–30µg 10µg 9.5–12.5µg 6µg 6–7µg 15–20µg 10–30µg RNase Treatment Optional Optional Optional Optional Optional Optional Optional Required Required Required Required Required Required Optional Insects Sf9 cells 5 × 10 6 cells 16µg Required Plant Tissue Tomato Leaf 40mg 7–12µg Required Gram Negative Bacteria Escherichia coli JM109 overnight culture, ~2 × 10 9 cells/ml Enterobacter cloacae overnight culture, ~6 × 10 9 cells/ml Gram Positive Bacteria Staphylococcus epidermis overnight culture, ~3.5 × 10 8 cells/ml Yeast Saccharomyces cerevisiae overnight culture, ~1.9 × 10 8 cells/ml 1ml 5ml 1ml 5ml 20µg 75–100µg 20µg 75–100µg Required Required Required Required 1ml 6–13µg Required 1ml 4.5–6.5µg Required Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.com Part# TM050 Printed in USA. Page 4 Revised 12/10
Page 1 and 2: Technical Manual Wizard ® Genomic
Page 3: included at this time; it is option
Page 7 and 8: ! If blood sample has been frozen,
Page 9 and 10: 4. Remove and discard as much super
Page 11 and 12: 5. Carefully remove and discard as
Page 13 and 14: . Add 10-20mg of fresh or thawed ti
Page 15 and 16: 3.F. 7. Carefully remove the supern
Page 17 and 18: 3.G. Isolating Genomic DNA from Gra
Page 19 and 20: 4. Troubleshooting (continued) Symp
Page 21: © 2010 Promega Corporation. All Ri

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