ISOLATION OF HIGH-MOLECULAR-WEIGHT DNA FROM MAMMALIAN-CELLS
ISOLATION OF HIGH-MOLECULAR-WEIGHT DNA FROM MAMMALIAN-CELLS
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DOI：
10.1111/j.1432-1033.1973.tb02881.x
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发表时间：
1973-01-01
期刊：
影响因子：
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通讯作者：
CHAMBON, P
中科院分区：
文献类型：
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作者：
GROSS-BELLARD, MM;OUDET, P;CHAMBON, P
描述了一种从动物细胞中分离高分子量DNA的制备方法。该方法基于蛋白酶K的使用，蛋白酶K是一种具有广泛作用谱的强效蛋白水解酶，在十二烷基硫酸钠和乙二胺四乙酸存在下非常活跃。DNA制备物不含RNA、蛋白质和降解酶。天然DNA的数均分子量为190 × 106，而单链DNA的数均分子量为90 × 106，表明DNA分子不含单链切口。天然DNA分子的分子量范围从40 × 106到500 × 106以上。
A preparative method for isolating high‐molecular‐weight DNA from animal cells is described. This method is based on the use of proteinase K, a powerful proteolytic enzyme with a broad action spectrum, which is very active in the presence of sodium dodecylsulfate and ethylene‐diamine tetraacetate. The DNA preparation is free of RNA, protein and degrading enzymes. The number‐average molecular weight of the native DNA is 190 × 106, whereas it is 90 × 106for single‐stranded DNA, indicating that the DNA molecules do not contain single‐stranded nicks. The native DNA molecules range in molecular weight from 40 × 106to more than 500 × 106.