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Northern Blotting Technique for Detection and Expression Analysis of mRNAs and Small RNAs

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RNA Abundance Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2170))

Abstract

Northern analysis is a conventional but gold standard method for detection and quantification of gene expression changes. It not only detects the presence of a transcript but also indicates size and relative comparison of transcript abundance on a single membrane. In recent years it has been aptly adapted to validate and study the size and expression of small noncoding RNAs. Here, we describe protocols employed in our laboratory for conventional northern analysis with total RNA/mRNA to study gene expression and validation of small noncoding RNAs using low molecular weight fraction of RNAs. A brief account on the recent advancements for improving the sensitivity and efficiency of northern blot detection is also included in this chapter.

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Acknowledgements

Research work in the laboratories of SK-A and MA is supported by funds from Department of Biotechnology (DBT), PURSE and FIST grants from Department of Science & Technology (DST), University Grants Commission (UGC) and University of Delhi. ARB and RP are thankful to Ramjas College, University of Delhi and SGTB Khalsa College, University of Delhi, respectively, for financial support. Ankur R. Bhardwaj and Ritu Pandey contributed equally to this work.

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Author notes

  1. Ankur R. Bhardwaj and Ritu Pandey contributed equally to this work.

Authors and Affiliations

  1. Department of Botany, Ramjas College, University of Delhi North Campus, Delhi, India

    Ankur R. Bhardwaj

  2. Department of Botany, SGTB Khalsa College, University of Delhi North Campus, Delhi, India

    Ritu Pandey

  3. Department of Botany, University of Delhi North Campus, Delhi, India

    Manu Agarwal

  4. Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India

    Surekha Katiyar-Agarwal

Authors
  1. Ankur R. Bhardwaj
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  2. Ritu Pandey
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  3. Manu Agarwal
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  4. Surekha Katiyar-Agarwal
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Corresponding author

Correspondence to Surekha Katiyar-Agarwal .

Editor information

Editors and Affiliations

  1. Department of Plant Pathology and Microbiology, Center for Plant Cell Biology and Institute for Integrative Genome Biology, University of California, Riverside, CA, USA

    Hailing Jin

  2. Department of Nematology, University of California, Riverside, CA, USA

    Isgouhi Kaloshian

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Bhardwaj, A.R., Pandey, R., Agarwal, M., Katiyar-Agarwal, S. (2021). Northern Blotting Technique for Detection and Expression Analysis of mRNAs and Small RNAs. In: Jin, H., Kaloshian, I. (eds) RNA Abundance Analysis . Methods in Molecular Biology, vol 2170. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0743-5_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0743-5_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0742-8

  • Online ISBN: 978-1-0716-0743-5

  • eBook Packages: Springer Protocols

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