Protocol for Genomic DNA Isolation from Mouse Tail/Animal Tissue or Cultured Cells
- Document # 27177
- Version 1.0.0
- Dec 2019
The following protocol is for genomic DNA isolation from cultured cells or animal tissue using the Genomic DNA Purification Kit (Catalog #79020). For complete instructions, refer to the Technical Manual (Document #10000005432).
Directions
A. Preparation of Cell Lysate
Prepare cell lysate from mouse tail or tissue, or from tissue culture cells, as indicated below.

Mouse Tail or Animal Tissue Lysate
- Prepare Digestion Solution as indicated in Table 1. Mix thoroughly and store on ice.
- Cut a 0.5 - 1.2 cm length of mouse tail from the tip or weigh up to 20 mg of tissue sample in a clean DNase-free 1.7 mL microcentrifuge tube.
- Add 275 μL Digestion Solution to each tube.
- Incubate the sample tubes overnight (16 - 18 hours) in a 55ºC heating block or water bath.
- Add 250 μL Lysis Buffer to each sample. Vortex to mix.
- Proceed to DNA isolation.
Table 1. Preparation of Digestion Solution
Tissue Culture Cell Lysate from Cell Suspension
- Collect 1 x 104 to a maximum of 5 x 106 cells. Wash the cells once with D-PBS.
- Add 150 μL Lysis Buffer to the washed cells. Mix by pipetting up and down.
- Proceed to DNA isolation.
B. DNA Isolation
- Insert minicolumn into Collection Tube.
- Transfer lysate sample to the minicolumn assembly.
- Centrifuge at 13,000 x g for 3 minutes. Remove the minicolumn from the Collection Tube and discard the liquid. Reinsert the minicolumn in the Collection Tube.
- Add 650 μL Column Wash Buffer (with ethanol added). Centrifuge at 13,000 x g for 1 minute. Remove the minicolumn from the Collection Tube and discard the liquid. Reinsert the minicolumn in the Collection Tube.
- Repeat step 5 for a total of 4 washes.
- Empty the Collection Tube and place the minicolumn back in the tube. Centrifuge at 13,000 x g for 2 minutes to dry the membrane.
- Carefully transfer minicolumn to a new labeled 1.7 mL microcentrifuge tube.
- Add 250 μL nuclease-free water to the minicolumn. Incubate at room temperature for 1 - 2 minutes. Centrifuge at 13,000 x g for 1 minute. For mouse tail or animal tissue lysates, proceed to step 9. For tissue culture lysates, proceed to step 10.
- Add an additional 250 μL nuclease-free water to the minicolumn. Incubate at room temperature for 1 - 2 minutes. Centrifuge at 13,000 x g for 1 minute.
- Discard minicolumn and store purified DNA at -20ºC.
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