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

From 2011.igem.org

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===Practice===
===Practice===
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10 mm^3
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1. Prepare small pieces of squid tissues
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1. Transfer embryo(s) into microfuge tube and remove excess liquid with drawn-out Pasteur pipette.
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2. Add extraction buffer, 10 mm^3 squid tissue with 100 µl PCR Extraction buffer in each tube, and incubate at 50-56°C for 2-3 hrs (longer is ok). Vortex occasionally.  
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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.  
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Revision as of 16:56, 19 July 2011

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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

1. Prepare small pieces of squid tissues

2. Add extraction buffer, 10 mm^3 squid tissue with 100 µl PCR Extraction buffer in each tube, 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.