DNA Extraction from Embryos for PCR

This method is quicker than the previous one and is suitable for embryos. The DNA is suitable for PCR, but is sheared so it is not suitable for Southern analysis.

1. Total nucleic acid is extracted from pools of 40 to several hundred dechorionated embryos at 16-24 h.

2. Add 10 vol of 4 M guanidinium isothiocyanate, 0.25 mM sodium citrate, pH 7.0, 0.5 % Sarkosyl, 0.1 M P-mercaptoethanol.

3. Vortex for 1 min.

4. Extract once with phenol:chloroform:isoamyl alcohol (25:24:1).

5. Precipitate nucleic acid by addition of 3 vol ethanol and 1/10 vol sodium acetate (3 M, pH 5.5).

6. Wash the pellet with 70% ethanol.

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