Efficientin vitrosynthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter

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ID: 289361
1984
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Abstract
A simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described. This invitro transcription system is based on the unusually specific RNA synthesis by bacteriophage SP6 RNA polymerase which initiates transcription exclusively at an SP6 promoter. We have constructed convenient cloning vectors that contain an SP6 promoter immediately upstream from a polylinker sequence. Using these SP6 vectors, optimal conditions have been established for invitro RNA synthesis. The advantages and uses of SP6 derived RNAs as probes for nucleic acid blot and solution hybridizations are demonstrated. We show that single stranded RNA probes of a high specific activity are easy to prepare and can significantly increase the sensitivity of nucleic acid hybridization methods. Furthermore, the SP6 transcription system can be used to prepare RNA substrates for studies on RNA processing (1,5,9) and translation (see accompanying paper).
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openalex_W2078119356 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Douglas A. Melton, Paul A. Krieg, Michael R. Rebagliati, Tom Maniatis, Kai Zinn, M. R. Green
Journal Nucleic Acids Research
Year 1984
DOI
10.1093/nar/12.18.7035
URL
https://academic.oup.com/nar/article-pdf/12/18/7035/6197415/12-18-7035.pdf
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