Solution structural studies of chromatin fibers.
Solution structural studies of chromatin fibers.
复制标题
染色质纤维的溶液结构研究。
DOI：
10.1021/bi00509a006
复制
发表时间：
1981
期刊：
影响因子：
2.9
通讯作者：
Crothers,DM
中科院分区：
文献类型：
--
作者：
Lee,KS;Mandelkern,M;Crothers,DM
核及溶解染色质的制备材料与方法。采用Blobel & Potter（1966）改良的方法制备小牛胸腺核。取50 g冷冻胸腺标本，在200 mL溶液A （0.3 M蔗糖、50 mM三乙醇胺、25 mM KC1、5 mM MgCl2和1 mM phch2so2f， pH 6.8, 4℃）中切碎，用aware搅拌器在40 V下搅拌5分钟。用粗棉布过滤后，将细胞核在800g下成球10分钟，先用溶液A，再用溶液B （0.3 M蔗糖，50 mM三乙醇胺，25 mM KC1, 4 mM MgCl2, 1 mM CaCl2和1 mM phch2so2f (PMSF)）洗涤数次。
Materials and MethodsPreparation of Nuclei and Solubilized Chromatin. Calf thymus nuclei were prepared by modification of the method of Blobel & Potter (1966). A 50-g sample of frozen thymus was minced and homogenized in 200 mL of solution A (0.3 M sucrose, 50 mM triethanolamine, 25 mM KC1, 5 mM MgCl2, and 1 mM PhCH2S02F, pH 6.8 at 4 C) by using aWaring blender at 40 V for 5 min. After filtration through cheesecloth, nuclei were pelleted at 800g for 10 min and washed several times with solution A and then with solution B (0.3 M sucrose, 50 mM triethanolamine, 25 mM KC1, 4 mM MgCl2, 1 mM CaCl2, and 1 mM PhCH2S02F (PMSF),