Sonifier W-450D, Branson

Материал из Zbio

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NB! pre-PCR operation

  1. dilute 1ng-1µg of DNA in 0.4-1ml of degased 1x TE;
  2. put DNA in 2ml non-autoclaved tube, cool it on ice;
  3. bring to the sonifier:
    • 1L mQ water
    • 700ml glass beaker
    • yellow tips
    • parafilm pieces
    • paper towels and lens cleaning tissue
    • ice
  4. wash the sonotrode two times in ~300ml mQ (40% amplitude, 10sec);
  5. dry the sonotrode (40% amplitude, 10sec), remove the rest of water with lens cleaning tissue;
  6. open the tube with DNA, cover it with parafilm and make a hole with a yellow tip;
  7. insert the sonotrode to about 1/2 level of the solution / make sure the sonotrode does not touch the walls;
  8. plunge the tube into the beaker with water/ice;
  9. 10 min of sonication (10% amplitude; sonication/cooling = 10sec/50sec; program 6; total time is ~1h);
  10. remove the tube with DNA, snap freeze in liquid nitrogen, store at -20°C;
  11. wash the sonotrode again.

[править] Notes

Sonotrode washing

  • routine wash:
» rinse with Bidest water
» 2x sonicate in glass beaker with mQ water
  • thorough wash:
» incubate 30min in 1% Na hypochorite
» wash thoroughly in water
» 2x sonicate in glass beaker with mQ water

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