Team:Cambridge/Protocols/Extraction of genomic DNA from squid

Extraction of genomic DNA from squid

A method to extract genomic DNA from squid tissue.

Large sample number, very quick and dirty, adequate for PCR

protocol adapted from http://zfin.org/zf_info/zfbook/chapt9/9.3.html

Theory

How it works

Practice

10 mm^3

1. Transfer embryo(s) into microfuge tube and remove excess liquid with drawn-out Pasteur pipette.

2. Add extraction buffer, 50 µl or 10 µl per embryo whichever is larger, and incubate at 50-56°C for 2-3 hrs (longer is ok). Vortex occasionally.

PCR Extraction buffer

10 mM Tris pH 8 2 mM EDTA 0.2% Triton X-100 200 µg/ml Proteinase K

3. Boil samples in waterbath for 5-10 min to inactivate Proteinase K.

4. Spin in microfuge for 1 min, store at - 20°C.

5. Use 10-15 µl to set up PCR reaction in a total volume of 50 µl, proceed with your favorite PCR protocol.

Safety

The safety implication of the procedure.