protocols

				Protocols


Services & Fees Mouse Colony Information Links Contact Us Home		Tail DNA Extraction 0.5-1 cm tail digested in 500 ul SE buffer/proteinase K, 55°C overnight. Add prewarmed saturated NaCl solution(6M) 170 ul, gently mix to a final concentration of 1.5 M Add 670 ul of chloroform and mix for 30-60 min by gentle rotation Centrifugate at 12000 rpm for 5min. Transfer the supernatant to a fresh tube,add 1 volume of isopropanol at room temperature and gently mix for 5 min. Pick DNA or centrifugate to collect DNA Wash with 70% ethanol for 1 hour or overnight. Spin and dry Dissolve in Tris 10 mM, pH8.5 SE buffer/Proteinase K

					Final concentration	Stock solution	Stock solutions needed to make 10 ml SE buffer
			NaCl		75 mM	5 M	150 µl
			EDTA pH8.0		25 mM	0.5 M	500 µl
			SDS		1%	10%	1000 µl
			H2O				8150 µl
			Proteinase K		200 µg/ml	10 mg/ml	200 µl
	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.