Isolation of Mouse Tail DNA for Screening

1. Cut 1-1.5 cm of tail from the mouse and mince with scissors in 400 pL of TE-SDS in a large Eppendorf tube.

2. Add proteinase K (5 mL; 20 pg/mL) to each sample, and incubate at 65°C overnight.

3. Add KAc (75 pL; 8 M) and chloroform (500 pL) to each sample, and mix by inversion.

4. Centrifuge for 5 min at 14,000g, collect the (upper) aqueous phase into an Eppendorf tube containing 1 mL ethanol (100%), and leave at room temperature for 5-10 min.

5. Spool precipitated DNA onto a sealed pipet, wash by dipping each sample in ethanol (90%) and air-dry (5-10 min).

6. Shake DNA off the pipet in 200 |L of TE in an Epppendorf tube and allow to dissolve overnight before calculating concentration by absorbence at 260 nm.

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