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Identification of Newly Transcribed RNA
Abstract
Newly transcribed RNA can be identified using the nuclear runoff transcription assay. In this assay, isolated nuclei, free of membranes and cytoplasmic debris, are used in an in vitro transcription reaction in the presence of 32 P‐labeled UTP. The labeled RNA can then be hybridized to cDNAs immobilized on nitrocellulose. This unit describes three methods for isolating suitable nuclei by detergent lysis, Dounce homogenization, and centrifugation on a sucrose gradient. Two Support Protocols describe the preparation of nitrocellulose filter strips containing double‐stranded and single‐stranded DNAs, which are used to detect the presence of specific transcripts in the nuclear runoff transcription assay.
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Citations
Cited by
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Iron regulates nitric oxide synthase activity by controlling nuclear transcription.
Weiss, G., Werner-Felmayer, G., Werner, E. R., Grünewald, K., Wachter, H., & Hentze, M. W. (1994) · The Journal of experimental medicine · doi:10.1084/jem.180.3.969 · Cited by 412
NF-κB Regulates Phagocytic NADPH Oxidase by Inducing the Expression of gp91
Anrather, J., Racchumi, G., & Iadecola, C. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m506172200 · Cited by 386
The splicing factor SC35 has an active role in transcriptional elongation
Lin, S., Coutinho-Mansfield, G., Wang, D., Pandit, S., & Fu, X.-D. (2008) · Nature Structural & Molecular Biology · doi:10.1038/nsmb.1461 · Cited by 362
Requirement for p38α in Erythropoietin Expression
Tamura, K., Sudo, T., Senftleben, U., Dadak, A. M., Johnson, R., & Karin, M. (2000) · Cell · doi:10.1016/s0092-8674(00)00027-1 · Cited by 340
Metadata
- Authors
- Greenberg, M. E., & Bender, T. P.
- Venue
- Current Protocols in Molecular Biology
- Publisher
- Wiley
- Published
- 2007-04
- Volume
- 78
- Issue
- 1
- Pages
- Unit 4.10-Unit 4.10
- ISSN
- 1934-3639 (ISSN-L)
- 1934-3647
- OpenAlex
- W1530430636
- Cited by
- 185
- References
- 0
- FWCI
- 1.68
- Type
- journal article
- Language
- en
- Open access
- closed
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- Greenberg, M. E., & Bender, T. P. (2007). Identification of Newly Transcribed RNA. Current Protocols in Molecular Biology. https://doi.org/10.1002/0471142727.mb0410s78
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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