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Identification and comprehensive evaluation of reference genes for RT-qPCR analysis of host gene-expression in Brassica juncea-aphid interaction using microarray data

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Title Identification and comprehensive evaluation of reference genes for RT-qPCR analysis of host gene-expression in Brassica juncea-aphid interaction using microarray data
Not Available
 
Creator Chet Ram
Murali Krishna Koramutla
Ramcharan Bhattacharya
 
Subject Aphid infestation
Arabidopsis
Brassica juncea
Normalization
RT-qPCR
Reference genes
 
Description Not Available
Brassica juncea is a chief oil yielding crop in many parts of the world including India. With advancement
of molecular techniques, RT-qPCR based study of gene-expression has become an integral part of experimentations
in crop breeding. In RT-qPCR, use of appropriate reference gene(s) is pivotal. The virtue of
the reference genes, being constant in expression throughout the experimental treatments, needs to be
validated case by case. Appropriate reference gene(s) for normalization of gene-expression data in
B. juncea during the biotic stress of aphid infestation is not known. In the present investigation, 11
reference genes identified from microarray database of Arabidopsis-aphid interaction at a cut off FDR
0.1, along with two known reference genes of B. juncea, were analyzed for their expression stability
upon aphid infestation. These included 6 frequently used and 5 newly identified reference genes.
Ranking orders of the reference genes in terms of expression stability were calculated using advanced
statistical approaches such as geNorm, NormFinder, delta Ct and BestKeeper. The analysis suggested CAC,
TUA and DUF179 as the most suitable reference genes. Further, normalization of the gene-expression data
of STP4 and PR1 by the most and the least stable reference gene, respectively has demonstrated
importance and applicability of the recommended reference genes in aphid infested samples of B. juncea.
Not Available
 
Date 2019-04-08T07:46:45Z
2019-04-08T07:46:45Z
2017-05-11
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/18136
 
Language English
 
Relation Not Available;
 
Publisher ELSEVIER