Mapping the genetic organization of RNA by electron microscopy.

摘要: Publisher Summary Electron microscopic (EM) analysis of DNA–DNA heteroduplexes permits the evaluation sequence and organizational relationships among related genomic subgenomic molecules. Formamide is used to dissociate adventitious intrastrand pairing during EM grid preparation, facilitating measurement both double-stranded single-stranded DNA segments. There are two forms RNA–DNA heteroduplexes. In an R-loop, remains duplexed, except where RNA has hybridized caused displacement its counterpart, leading formation a loop. RNA-single-stranded (R–D heteroduplexes), unpaired, hybridized. The composition hybridization solution narrowly dictated by high concentration formamide required. NaCl should be as practicable promote maximal rate annealing.