Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)
Identifieur interne : 000A94 ( Main/Exploration ); précédent : 000A93; suivant : 000A95Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)
Auteurs : Neha Kapila [Inde] ; Amit Kishore [Inde] ; Monika Sodhi [Inde] ; Ankita Sharma [Inde] ; Pawan Kumar [Inde] ; A. K. Mohanty [Inde] ; Tanushri Jerath [Inde] ; M. Mukesh [Inde]Source :
- ISRN Biotechnology [ 2090-9403 ] ; 2013.
Abstract
Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified
Url:
DOI: 10.5402/2013/735053
PubMed: 25937980
PubMed Central: 4393032
Affiliations:
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K." last="Mohanty">A. K. Mohanty</name><affiliation wicri:level="1"><nlm:aff id="I3">Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Jerath, Tanushri" sort="Jerath, Tanushri" uniqKey="Jerath T" first="Tanushri" last="Jerath">Tanushri Jerath</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Mukesh, M" sort="Mukesh, M" uniqKey="Mukesh M" first="M." last="Mukesh">M. Mukesh</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author></analytic><series><title level="j">ISRN Biotechnology</title><idno type="eISSN">2090-9403</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified <italic>RPL4, EEF1A1,</italic> and <italic>RPS23</italic> genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Marai, I F M" uniqKey="Marai I">I. F. M. Marai</name></author><author><name sortKey="Haeeb, A A M" uniqKey="Haeeb A">A. A. M. 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Glare</name></author><author><name sortKey="Divjak, M" uniqKey="Divjak M">M. Divjak</name></author><author><name sortKey="Bailey, M J" uniqKey="Bailey M">M. J. Bailey</name></author><author><name sortKey="Walters, E H" uniqKey="Walters E">E. H. Walters</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bower, N I" uniqKey="Bower N">N. I. Bower</name></author><author><name sortKey="Johnston, I A" uniqKey="Johnston I">I. A. Johnston</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Kapila, Neha" sort="Kapila, Neha" uniqKey="Kapila N" first="Neha" last="Kapila">Neha Kapila</name></noRegion><name sortKey="Jerath, Tanushri" sort="Jerath, Tanushri" uniqKey="Jerath T" first="Tanushri" last="Jerath">Tanushri Jerath</name><name sortKey="Kapila, Neha" sort="Kapila, Neha" uniqKey="Kapila N" first="Neha" last="Kapila">Neha Kapila</name><name sortKey="Kishore, Amit" sort="Kishore, Amit" uniqKey="Kishore A" first="Amit" last="Kishore">Amit Kishore</name><name sortKey="Kumar, Pawan" sort="Kumar, Pawan" uniqKey="Kumar P" first="Pawan" last="Kumar">Pawan Kumar</name><name sortKey="Mohanty, A K" sort="Mohanty, A K" uniqKey="Mohanty A" first="A. K." last="Mohanty">A. K. Mohanty</name><name sortKey="Mukesh, M" sort="Mukesh, M" uniqKey="Mukesh M" first="M." last="Mukesh">M. Mukesh</name><name sortKey="Sharma, Ankita" sort="Sharma, Ankita" uniqKey="Sharma A" first="Ankita" last="Sharma">Ankita Sharma</name><name sortKey="Sodhi, Monika" sort="Sodhi, Monika" uniqKey="Sodhi M" first="Monika" last="Sodhi">Monika Sodhi</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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