Koller B, Delius H, Bünemann H, Müller W
Gene. 1978 Nov;4(3):227-39. doi: 10.1016/0378-1119(78)90020-3.
Electrophoretic elution of DNA coupled with direct adsorption onto malachite green-polyacrylamide columns was used to isolate double- and single-stranded DNA from agarose gels. Subsequently, DNA was eluted with a high salt buffer and filtered through Sephadex which permitted recovery of the DNA in a low salt buffer at concentrations suitable for heteroduplex analysis by electron microscopy. This method was tested by examining heteroduplexes formed from the isolated complementary single strands of T7 wild type DNA and a T7 deletion mutant. More than 80% of the reannealed molecules were intact heteroduplexes showing the deletion loop. Irradiation of single-stranded DNA with 254 nm light resulted in distorted, convoluted heteroduplexes while 366 nm light did not show this effect.
将DNA的电泳洗脱与直接吸附到孔雀石绿-聚丙烯酰胺柱上相结合，用于从琼脂糖凝胶中分离双链和单链DNA。随后，用高盐缓冲液洗脱DNA，并通过葡聚糖凝胶过滤，从而在低盐缓冲液中以适合电子显微镜异源双链分析的浓度回收DNA。通过检查由T7野生型DNA和T7缺失突变体的分离互补单链形成的异源双链体来测试该方法。超过80%的重新退火分子是完整的显示缺失环的异源双链体。用254nm光照射单链DNA会导致异源双链体扭曲、盘绕，而366nm光则没有这种效果。
