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文献和实验Site-directed hydroxy radical mapping of nucleosome positions in vitro
all on a pre-run 160x200x1mm, 5% (1:40) acrylamide, 0.2xTBE native gel equilibrated at 4°C. The gel is both pre-run and run at 300V for 180-240 minutes. When the sample contains larger amounts of nucleosome it may be necessary to increase the amount of ferrous
RNA AND PROTEIN EXTRACTION FROM THE SAME SAMPLE
Tris-HCl pH 8; 1% SDS, 1% deoxycholate-sodium, 20 mM EDTA (2% PVP 40,000-optional when using tissues with high conconcentration of phenols) 2. Phenol: Add to 200 mL liquified phenol 200 mg of antioxidant
Purification of dnEBNA-1/Soft from E. coli BL21 LysS
Inoculate 2ml of 5ml o/n culture of either p3133 (empty vector pET11a)or p3134 (dnEBNA-1/Soft)in E.coli BL21 LysS per 0.5L LB+ ampicillin (grow two 0.5L cultures of each) Incubate ~2hrs 37℃250rpm until OD600 = 0.4-0.6 Induce with 5ml 100mM ITPG
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