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  • Whole mount in situ localization of miRNAs and target mRNA transcripts in plants.

Whole mount in situ localization of miRNAs and target mRNA transcripts in plants.

3 Biotech (2019-05-09)
Vibhav Gautam, Archita Singh, Swati Verma, Sharmila Singh, Sourav Chatterjee, Ananda K Sarkar
摘要

The functional characterization of miRNAs often involves understanding of their spatiotemporal expression, which mostly relies on reporter-based or in situ hybridization studies. The available in situ localization methods follow separate protocols for pre-hybridization, hybridization, post-hybridization, and detection steps for both miRNA and mRNA transcripts in plants. In this study, we present a single method which can be used for whole mount in situ localization of both miRNAs and mRNAs in different plant tissues. Our modified method provides enhanced sensitivity for the localization of miRNA and their target transcripts. Consequently, a less laborious, time-saving, economic and efficient method has been proposed by the modification of pre-hybridization, hybridization, post-hybridization and detection steps.

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Sigma-Aldrich
Trizma ® 碱, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
甘氨酸, suitable for electrophoresis, ≥99%
Sigma-Aldrich
氢氧化钠, BioXtra, ≥98% (acidimetric), pellets (anhydrous)
Sigma-Aldrich
HistoChoice ® 透明剂, alternative to toluene and xylene
Sigma-Aldrich
磷酸钠 一元, BioXtra, ≥99.0%