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Tail DNA Extraction

  • 0.5 to 1 cm tail digested in 500 ul SE buffer/proteinase K, 55C overnight.
  • Add pre-warmed saturated NaCl solution(6M) 170 ul, gently mix to a final concentration of 1.5 M.
  • Add 670 ul of chloroform and mix for 30 to 60 minutes by gentle rotation.
  • Centrifugate at 12000 rpm for five minutes.
  • Transfer the supernatant to a fresh tube, add one volume of isopropanol at room temperature and gently mix for five minutes.
  • Pick DNA or centrifugate to collect DNA.
  • Wash with 70 percent ethanol for one hour or overnight.
  • Spin and dry.
  • Dissolve in Tris 10 mM, pH8.5.

SE Buffer/Proteinase K

 
Final Concentration
Stock Concentration
For 10 ml Mix
NaCl 75 mM 5 M 150 l
EDTA pH8.0 25 mM 0.5 M 500 l
SDS 1% 10% 1000 l
H2O     8150 l
Proteinase K 200 g/ml 10 mg/ml 200 l

Reference

R. MGllenbach, PJL Lagoda and C Welter (1989). An Efficient Salt-Chloroform Extraction of DNA from blood and tissues. Trend in Genetics 5(12):391.