Goldbach R W, Evers R F, Borst P
Nucleic Acids Res. 1978 Aug;5(8):2743-54. doi: 10.1093/nar/5.8.2743.
We have found that binding of poly(U,G) to single-stranded DNA decreases its mobility in 0.3% agarose gels. Differential binding to the complementary strands of denatured duplex DNA provides a simple method for strand separation. The method is shown to work with bacteriophage lambda DNA, adenovirus DNA and mtDNA for Tetrahymena pyriformis. In all cases the strand that binds more poly(U,G) in CsCl gradients also binds more in gels. The separated strands can be directly blotted from the gel onto nitrocellulose filters and used for hybridization experiments.
我们发现,聚(U,G)与单链DNA的结合会降低其在0.3%琼脂糖凝胶中的迁移率。与变性双链DNA互补链的差异结合提供了一种简单的链分离方法。该方法已证明可用于噬菌体λDNA、腺病毒DNA和梨形四膜虫的线粒体DNA。在所有情况下,在CsCl梯度中结合更多聚(U,G)的链在凝胶中也结合更多。分离出的链可直接从凝胶转移到硝酸纤维素滤膜上,用于杂交实验。