IMPROVED METHODS FOR THE FORMATION AND STABILIZATION OF R-LOOPS

被引：153

作者：

KABACK, DB

ANGERER, LM

DAVIDSON, N

机构：

[1] Department of Chemistry, California Institute of Technology, Pasadena

来源：

NUCLEIC ACIDS RESEARCH | 1979年 / 6卷 / 07期

关键词：

D O I：

10.1093/nar/6.7.2499

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Improved methods for the formation and stabilization of R-loops for visualization in the electron microscope are presented. The two complementary strands of a duplex DNA are photochemically crosslinked once every 1 to 3 kb using 4, 5′, 8 trimethylpsoralen. R-loops are then formed by incubation with RNA in 70% formamide at a temperature above the DNA melting temperature. Finally, the R-loops are stabilized by modifying the free single strand of DNA with glyoxal, thus minimizing the displacement of the hybridized RNA by branch migration. In this manner R-loops can be formed and visualized at a high frequency irrespective of the base composition of the nucleic acid of interest. © 1979 Information Retrieval Limited.

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页码：2499 / 2517

页数：19

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