Detection of Small Products of Transcription from 6S RNA (pRNA) by 'Mirror-Like' Northern Blot Hybridization
| Autor: | Elena A. Kubareva, Olga Y. Burenina, T. S. Oretskaya |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Gel electrophoresis 010405 organic chemistry Chemistry Organic Chemistry RNA 01 natural sciences Biochemistry 0104 chemical sciences 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology Transcription (biology) Complementary DNA RNA polymerase microRNA Digoxigenin Northern blot |
| Zdroj: | Russian Journal of Bioorganic Chemistry. 47:478-482 |
| ISSN: | 1608-330X 1068-1620 |
| Popis: | Investigation of RNAs with a length less than 30 nucleotides, in particular, microRNAs and other extremely short noncoding RNAs, is often associated with difficulties in detecting them not only in vivo, but also in vitro. One of these molecules is bacterial pRNA (product RNA) synthesized by RNA polymerase on 6S RNA as a template. The classical method for identifying pRNA is Northern blot hybridization with complementary DNA probes containing various labels and modified nucleosides. However, this method is not suitable for searching for new pRNAs whose sequences are unknown. It is also quite expensive and time-consuming. We have proposed a new simple way to determine the fact of pRNA synthesis on the template of 6S RNA that we called “mirror-like” Northern blotting. It does not require gel electrophoresis, and the pRNA, newly formed during transcription, act as a probe itself due to incorporated digoxigenin (DIG) residues. This approach will allow fast screening of new 6S RNAs or their mutant variants for their ability to serve as templates for pRNA synthesis and the estimation of its efficacy. |
| Databáze: | OpenAIRE |
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