Wahl G M, Stern M, Stark G R
Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683-7. doi: 10.1073/pnas.76.8.3683.
We describe a technique for transferring electrophoretically separated bands of double-stranded DNA from agarose gels to diazobenzyloxymethyl-paper. Controlled cleavage of the DNA in situ by sequential treatment with dilute acid, which causes partial depurination, and dilute alkali, which causes cleavage and separation of the strands, allows the DNA to leave the gel rapidly and completely, with an efficiency independent of its size. Covalent attachment of DNA to paper prevents losses during subsequent hybridization and washing steps and allows a single paper to be reused many times. Ten percent dextran sulfate, originally found to accelerate DNA hybridization in solution by about 10-fold [J.G. Wetmur (1975) Biopolymers 14, 2517-2524], accelerates the rate of hybridization of randomly cleaved double-stranded DNA probes to immobilized nucleic acids by as much as 100-fold, without increasing the background significantly.
我们描述了一种将琼脂糖凝胶中双链DNA的电泳分离条带转移至重氮苄氧基甲基纸的技术。通过用稀酸（导致部分脱嘌呤）和稀碱（导致链的切割和分离）顺序处理，在原位对DNA进行可控切割，使DNA能快速、完全地离开凝胶，其效率与DNA大小无关。DNA与纸的共价连接可防止在后续杂交和洗涤步骤中出现损失，并允许同一张纸多次重复使用。最初发现10%的硫酸葡聚糖能使溶液中的DNA杂交速度加快约10倍[J.G. 韦特穆尔（1975年）《生物聚合物》14卷，2517 - 2524页]，它能使随机切割的双链DNA探针与固定化核酸的杂交速度加快多达100倍，且不会显著增加背景。
