BENDICH A, BORENFREUND E, HUTCHISON D J
J Cell Biol. 1963 Oct;19(1):59-69. doi: 10.1083/jcb.19.1.59.
A method for the isolation of metaphase chromosomes from mouse L1210 leukemia cells has been developed. Cells, arrested at metaphase with colchicine, were exposed to hypotonic solution and the pH was then adjusted to 5.6 to stabilize the chromosomes. The metaphase figures were subsequently disrupted and the chromosomes isolated by a series of differential centrifugations in sucrose. The isolated chromosomes were well preserved, as judged by morphological criteria. The effect of various enzymes and chemical agents on the isolated chromosomes was studied. Chymotrypsin, trypsin, and deoxyribonuclease caused a marked disintegration of the chromosomes, whereas treatment with pepsin and ribonuclease induced no significant morphological alterations.
已开发出一种从小鼠L1210白血病细胞中分离中期染色体的方法。用秋水仙碱使细胞停滞在中期，将其置于低渗溶液中，然后将pH值调至5.6以稳定染色体。随后破坏中期图像，并通过在蔗糖中进行一系列差速离心来分离染色体。根据形态学标准判断，分离出的染色体保存良好。研究了各种酶和化学试剂对分离出的染色体的影响。胰凝乳蛋白酶、胰蛋白酶和脱氧核糖核酸酶导致染色体明显解体，而用胃蛋白酶和核糖核酸酶处理未引起明显的形态学改变。
