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ES Cell DNA Extraction from 96 Well Plates

  • Remove media with multiple chanel pipette or by aspiration.
  • Add 100 l PBS, remove.
  • Add 50 l lysis buffer.
  • Wrap the plate with wet paper towel then seal it in a plastic bag.
  • Incubate at 50C overnight.
  • Add 100 l cold 100 percent etOH.
  • Leave at room temp (not shaking).
  • If a plate spinner is available, spin plates for 5’ at 1000 rpm at 10C. Pour off EtOH by blotting on paper towel pad.
  • Wash three times with 150 l 70 percent EtOH and pour off.
  • Pat on paper towel pad until very little liquid comes out

Lysis Buffer

Final Concentration
Stock Solution
10 ml Lysis Buffer
0.5% Sarcosyl (Sigma L9150) 10% 500 l
200 mM NaCl 5 M 400 l
10 mM EDTA (pH 8.0) 0.5 M 200 l
10 mM Tris.HCl (pH 8.0) 1 M 100 l
1 mg/ml proteinase K 20 mg 500 l