一种制备完整核DNA的通用方法。

EMBO J. 1984 Aug;3(8):1837-42. doi: 10.1002/j.1460-2075.1984.tb02056.x.

Naked nuclear DNA is easily sheared. Two general methods are described for preparing intact DNA in a stable form that can be pipetted without breaking it. Cells are encapsulated in agarose microbeads and then lysed in a non-ionic detergent (i.e., Triton X-100) and 2 M NaCl or an ionic detergent (e.g., sodium or lithium dodecyl sulphate) in low salt. Most cellular protein and RNA then diffuse out through pores in the beads to leave encapsulated and naked DNA which is nevertheless accessible to enzymes and other probes. Remarkably, considerable structure is preserved since the DNA is supercoiled and chromosomes retain their shape.

裸露的核DNA很容易被剪切。本文描述了两种一般方法，用于制备稳定形式的完整DNA，这种DNA可以用移液器操作而不会断裂。细胞被包裹在琼脂糖微珠中，然后在低盐条件下用非离子去污剂（即 Triton X-100）和2 M NaCl或离子去污剂（例如十二烷基硫酸钠或锂盐）进行裂解。然后，大多数细胞蛋白质和RNA通过微珠上的孔扩散出去，留下被包裹的裸露DNA，而酶和其他探针仍可接触到这些DNA。值得注意的是，由于DNA是超螺旋的，染色体保留了它们的形状，因此相当多的结构得以保留。