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Team:Cambridge/Protocols/Extraction of genomic DNA from squid
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1. Transfer embryo(s) into microfuge tube and remove excess liquid with drawn-out Pasteur pipette. | 1. Transfer embryo(s) into microfuge tube and remove excess liquid with drawn-out Pasteur pipette. | ||
Revision as of 16:43, 19 July 2011
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.
