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

From 2011.igem.org

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PCR Extraction buffer
PCR Extraction buffer
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10 mM Tris pH 8
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*10 mM Tris pH 8
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2 mM EDTA
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*2 mM EDTA
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0.2% Triton X-100
+
*0.2% Triton X-100
-
200 µg/ml Proteinase K
+
*200 µg/ml Proteinase K

Revision as of 16:59, 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.