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Improved methods for the formation and stabilization of R-loops
Open accessAbstract
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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Cited by
Characterization of the glycinin gene family in soybean.
Nielsen, N. C., Dickinson, C. D., Cho, T. J., Thanh, V. H., Scallon, B. J., Fischer, R. L., Sims, T. L., Drews, G. N., & Goldberg, R. B. (1989) · The Plant Cell · doi:10.1105/tpc.1.3.313 · Cited by 382
High-efficiency prime editing with optimized, paired pegRNAs in plants
Lin, Q., Jin, S., Zong, Y., Yu, H., Zhu, Z., Liu, G., Kou, L., Wang, Y., Qiu, J.-L., Li, J., et al. (2021) · Nature Biotechnology · doi:10.1038/s41587-021-00868-w · Cited by 325
From DNA sequence to biological function
Oliver, S. G. (1996) · Nature · doi:10.1038/379597a0 · Cited by 310
Metadata
- Authors
- Kaback, D. B., Angerer, L. M., & Davidson, N.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1979
- Volume
- 6
- Issue
- 7
- Pages
- 2499-2517
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W1968121029
- Cited by
- 160
- References
- 0
- FWCI
- 10.38
- Type
- journal article
- Language
- en
- Open access
- green
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- Kaback, D. B., Angerer, L. M., & Davidson, N. (1979). Improved methods for the formation and stabilization of R-loops. Nucleic Acids Research. https://doi.org/10.1093/nar/6.7.2499
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