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Team:Cambridge/Protocols/Extraction of genomic DNA from squid
From 2011.igem.org
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| - | PCR Extraction buffer | + | :PCR Extraction buffer |
| - | *10 mM Tris pH 8 | + | ::*10 mM Tris pH 8 |
| - | *2 mM EDTA | + | ::*2 mM EDTA |
| - | *0.2% Triton X-100 | + | ::*0.2% Triton X-100 |
| - | *200 µg/ml Proteinase K | + | ::*200 µg/ml Proteinase K |
Revision as of 17:00, 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.
