Freeze-thaw method for recovering DNA fragments

1. Carefully cut the strip containing the DNA to be recovered under the UV lamp, smash the cut strip (less than 0.6g) and place it in a 1.5ml centrifuge tube;

2. Add an equal volume of Tris-HCl saturated phenol (pH 7.6) and mix by shaking;

3. Place at -20 °C for 5-10 min;

4. Centrifuge at 10000g×5min at 4°C, transfer the upper layer to another centrifuge tube;

5. Add 1/4 volume of H2O to the gel-containing centrifuge tube and mix by shaking;

6. -20 ° C, placed 5-10min;

7. Centrifuge at 10000g × 5min at 4 ° C, and combine the supernatant;

8. Extract once with an equal volume of phenol/chloroform or chloroform and take the supernatant;

9. Add 1/10 volume of 3M NaAc (pH 5.2), 2.5 volumes of pre-cooled absolute ethanol, and mix;

10. -20 ° C, let stand for 30 min;

11. Centrifuge 13000g×10min at 4°C, discard the supernatant, wash the precipitate with 5% times with 75% ethanol, and dry it;

12. Add an appropriate amount of H2O or TE to dissolve the precipitate.

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