Isolation of Genomic DNA from Mouse Tails

JoVE Journal
Biology

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JoVE Journal Biology

Isolation of Genomic DNA from Mouse Tails

Cut tail pieces (3mm) and mark ears.
Add 720 ml STE and 30 ml Proteinase K (10 mg/ml stock).
Incubate at 55°C on heating block and vortex every hour for 3 hours at top speed for 10 seconds.
Inactivate proteinase K at 70°C for 5 minutes.
Quench on ice for 5 minutes.
Centrifuge tail DNA for 10 minutes at full speed.
Decant into new tube containing 720 ml Isopropanol.
Precipitate DNA by inverting the tube or vortexing.
Spin down DNA for 5 minutes at full speed. Remove supernatant.
Wash pellet with 70% ethanol.
Spin down genomic DNA 5 minutes at full speed. Remove supernatant.
Allow DNA to dry for 1-2 minutes.
Resuspend DNA in 100-200 ml depending on size of pellet.
Place tube at 55°C for 1 hour to facilitate dissolution of DNA.

Learning Objectives

Material Name	Type	Company	Catalogue Number	Comment
STE	Buffer			100 mm Tris; pH 8.5 / 5 mM EDTA / 0.2% SDS / 200 mM NaCl / in H2O
TE	Buffer			100 mM Tris; pH 5.0 / 1 mM EDTA / 10 mM NaCl