Squish Buffer DNA extraction

Procedure

1. Homogenize tissue in 50 µl of squish buffer.

2. Add and additional 100 µl of squish buffer.

3. Add 1 µl of Proteinase K (4 µg/ml).

4. Incubate 60 min at 37° C; increase temperature to 85° C and incubate for 10 min.

5. Use 1-2 µl of DNA extraction for PCR.

Materials necessary

1.5 ml microcentrifuge pestle tubes
Mini pestles for 1.5ml tubes
Squish buffer
Proteinase K
10, 20 and 200 µl pipette tips

Equipment necessary

37° C incubator
85° C incubator
10 µl micropipette
20 µl micropipette
200 µl micropipette

Note: Squish buffer extractions do not last as long as other methods and should be stored at -20° C for no more than 30 days.

Adapted from the method of Gloor et al. 1991. Targeted gene replacement in Drosophila via P element-induced gap repair. Science 253: 1110. 

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