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VP 4 gene specific RT-PCR for detection of bovine group A rotaviruses

Indian Agricultural Research Journals

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Title VP 4 gene specific RT-PCR for detection of bovine group A rotaviruses
 
Creator MINAKSHI, MINAKSHI
MALIK, Y
PRASAD, G
PANDEY, R
 
Subject Bovine rotaviruses detection, Genome segment 4, RNA-PAGE, RT-PCR
 
Description RT-PCR assay was standardised with partial length primers specific to VP4 gene of bovine group A rotaviruses. The double stranded RNA (dsRNA) was extracted from MA 104 cell culture grown prototype rotaviruses by GIT lysis method. The presence of dsRNA in the extracts was confirmed by RNA-polyacrylamide gel electrophoresis (RNA-PAGE) with silver staining. To develop the RT-PCR assay dsRNA was first converted into complimentary DNA (cDNA) by reverse transcriptase enzyme and subsequently amplified by Taq DNA polymerase. The oligonucleotide primers used in RT-PCR assay were specific to both 3' and 5' ends of VP4 gene, which is highly, conserved among group A rotaviruses, The presence of expected 876 bp long PCR product was visualised by agarose gel electrophoresis. The primer pair used in present study specifically amplified VP4 gene. The results of present study suggested that RT -PCR assay could be used for detection of rotaviruses.
 
Publisher Directorate of Knowledge Management in Agriculture
 
Contributor —
 
Date 2014-01-21
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
—
 
Format application/pdf
 
Identifier http://epubs.icar.org.in/ejournal/index.php/IJAnS/article/view/36788
 
Source The Indian Journal of Animal Sciences; Vol 71, No 7 (2001)
0367-8318
 
Language eng
 
Relation http://epubs.icar.org.in/ejournal/index.php/IJAnS/article/view/36788/16407
 
Rights Copyright (c) 2014 The Indian Journal of Animal Sciences