The frozen tissue was sliced and then homogenized at 20 degree C in LiCl, 2 M; lauryl-trimethyl ammonium chloride (K & K No. 4484), 5%; pronase, B grade (calbiochem), 1 mg/ml and Tris-HCl 10 mM pH 7.5. The homogenate was left to stand 15 min at 20 degree C with occasional shaking. After centrifugation at 35 000 X g for 30 min at 0 degree C, the supernatant containing the crude DNA was purified by filtration on Ultrogel A 2 (LKB, Sweden). The Ultrogel A 2 column (2.5 X 45 cm) was equilibrated with a solution containing NaCl 2 M, EDTA 2.5 mM, Tris-HCl 10 mM pH 7.5. The flow rate was 3 ml cm-2 h-1. Five ml of the supernatant were placed on the column. The first peak contained highly polymerized (as demonstrated by ultracentrifugation) pure DNA (A260/A230 = 3.19; A260/A280 = 1.82). The yield was 2.26 mg of DNA/g of fresh liver.

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http://dx.doi.org/10.3109/13813458109069464DOI Listing

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