Identification of suitable housekeeping genes for normalization of quantitative real‐time PCR data during different physiological stages of mammary gland in riverine buffaloes (Bubalus bubalis). Issue 6 (30th January 2013)

• Record Type: Journal Article
• Authors: Aggarwal, J.; Sharma, A.; Kishore, A.; Mishra, B. P.; Yadav, A.; Mohanty, A.; Sodhi, M.; Kataria, R. S.; Malakar, D.; Mukesh, M.
• Is Part Of: Journal of animal physiology and animal nutrition. Volume 97:Issue 6(2013:Dec.)
• Page Start: 1132
• Page End: 1141
• Publication Date: 2013-01-30
• Subjects: Animal nutrition -- Periodicals; Feeds -- Periodicals; 636.085
• DOI: 10.1111/jpn.12027
• Languages: English
• ISSNs: 0931-2439

Abstract

<abstract abstract-type="main" id="jpn12027-abs-0001"> <title>Summary</title> <p>Gene expression analysis unravels the complex changes or relations at transcriptomic level. To nullify all type of errors that can be incorporated during any stage of RNA extraction into cDNA synthesis and for reliable results, the data obtained from qPCR have to be normalized using the appropriate/suitable housekeeping genes (HKGs). Unfortunately, till date, no such HKG has been reported for bubaline mammary gland. The objective of the present study was thus to identify and validate the potential HKGs for the gene expression studies in buffalo mammary gland. Mammary tissues from twelve buffaloes during different physiological stages: pre‐pubertal (heifer), lactation and involution were obtained for the present study. A total of 16 potential HKGs (<italic>GAPDH, β‐actin, UXT, β2M, A2M, RPl4, RPS9, RPS15A, RPS18, RPS23, HMBS, HPRT1, GTP, EEF1A1, UB1</italic> and <italic>RPL22</italic>) from different functional classes were evaluated. The analysis revealed that the expression of <italic>EEF1A1, RPl4, β2M</italic> and <italic>RPS15A</italic> was most consistent across different physiological stages of buffalo mammary gland. On the other hand<italic>, β‐actin</italic>, <italic> A2M</italic>, <italic> RPL22</italic> and <italic>GAPDH</italic> were the least stable genes making them unsuitable as HKGs. Based on our analysis, we recommend the use of <italic>EEF1A1, RPl4, β2M</italic> and <italic>RPS15A</italic> genes as suitable HKGs for accurate normalization of gene expression data in bubaline mammary gland.</p> </abstract>