Detection of small products of transcription from 6S RNA (pRNA) by “mirror-like” northern blot hybridization

Burenina, O.Y.; Oretskaya, T.S.; Kubareva, E.A.

Авторы: Burenina O.Y., Oretskaya T.S., Kubareva E.A.
Журнал: Russian Journal of Bioorganic Chemistry
Том: 47
Номер: 2
Год издания: 2021
Издательство: Maik Nauka/Interperiodica Publishing
Местоположение издательства: Russian Federation
Первая страница: 477
Последняя страница: 481
Оригинальные версии: Детекция малых пРНК – продуктов транскрипции 6S РНК – с помощью нозерн-блот-гибридизации в "зеркальном" варианте
DOI: 10.1134/S1068162021020060
Аннотация: 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 on6S RNA as a template. The classical method for identifying pRNA is Northern blot hybridization with complementaryDNA 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 “mirrorlike” Northern blotting. It does not require gel electrophoresis, and the pRNA,newly formed during transcription, act as a probe itself due to its containing 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.
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Detection of small products of transcription from 6S RNA (pRNA) by “mirror-like” northern blot hybridizationстатья

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