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. 1979 Jun 11;6(7):2499-317.
doi: 10.1093/nar/6.7.2499.

Improved methods for the formation and stabilization of R-loops

D B Kaback, L M Angerer, N Davidson
Free PMC article

Improved methods for the formation and stabilization of R-loops

D B Kaback et al. Nucleic Acids Res. .
Free PMC article

Abstract

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.

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References

    1. J Biol Chem. 1978 Nov 25;253(22):8239-46 - PubMed
    1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):725-9 - PubMed
    1. Biochemistry. 1978 Sep 5;17(18):3883-8 - PubMed
    1. Nucleic Acids Res. 1977;4(5):1539-52 - PubMed
    1. Proc Natl Acad Sci U S A. 1979 Feb;76(2):857-61 - PubMed

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