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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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Original Research Articles                      Volume : 9, Issue:2, February, 2020

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com /
submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2020.9(2): 215-221
DOI: https://doi.org/10.20546/ijcmas.2020.902.027


Chicken RNA Integrity Assessment through Capillary Electrophoresis for miRNA Expression Study using RT-qPCR
M. R. Vishnuraj1*, Suresh Devatkal1, S. Vaithiyanathan1, S. K. Mendiratta2, R. Uday Kumar1, Ch. Srinivas1 and M. Sowmya1
1ICAR-National Research Centre on Meat, Chengicherla, Hyderabad-92, Telangana, India
2ICAR-Indian Veterinary Research Institute, Izzatnagar, Bareilly, Uttarpradesh, India
*Corresponding author
Abstract:

The main concern in RNA research is the risk of degradation of RNA in the experimental conditions due to the ubiquitous nucleases and detecting the RNA quality through RNA integrity score provides better understanding of degradation process. Conventionally, the total RNA quantity and purity measured using spectrophotometer but even a standard reading stands questionable due to contributions from protein and genomic DNA and doesn’t ensure appropriate discrimination among RNA fractions. In present study, we performed RNA integrity analysis using capillary electrophoresis system for extracts from chicken liver, heart and gizzard. The optical measurement of RNA quality through measurement of A 260/280 ratio stands non reliable since below par RIN (RNA Integrity Number) were reported for both liver and gizzard extracts in capillary electrophoresis whereas heart extracts reported a RIN of 7.6 (optimum is 7). The RIN score of 6.8 for liver and 5.4 for gizzard were well correlated with the degraded size of 28S subunit and 18S subunit of ribosomal RNA respectively. In comparison with conventional method, capillary electrophoresis based RNA quality evaluation provides reliable information and such data can be used in selecting the RNA extracts for miRNA expression study in RT-qPCR.


Keywords: RNA, Degradation, RNA Integrity Score, Optical measurement, Capillary electrophoresis, Ribosomal RNA, Micro RNA

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How to cite this article:

Vishnuraj, M. R., Suresh Devatkal, S. Vaithiyanathan, S. K. Mendiratta, R. Uday Kumar, Ch. Srinivas and Sowmya, M. 2020. Chicken RNA Integrity Assessment through Capillary Electrophoresis for miRNA Expression Study using RT-qPCR.Int.J.Curr.Microbiol.App.Sci. 9(2): 215-221. doi: https://doi.org/10.20546/ijcmas.2020.902.027
Copyright: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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