• Users Online: 249
  • Print this page
  • Email this page
ORIGINAL ARTICLE
Year : 2020  |  Volume : 9  |  Issue : 5  |  Page : 247-254

Identification of stable internal control genes for accurate normalization of real-time quantitative PCR data in testicular tissue from two breeds of cattle


1 Theriogenology Laboratory, Veterinary Gynaecology, and Obstetrics, Southern Regional Station of ICAR- National Dairy Research Institute, Bengaluru – 560030 Karnataka, India
2 Animal Genomics Laboratory, ICAR - National Dairy Research Institute, Karnal – 132 001 Haryana, India
3 Southern Regional Station of ICAR-National Dairy Research Institute, Bengaluru – 560030 Karnataka, India

Correspondence Address:
Arumugam Kumaresan
Theriogenology Laboratory, Veterinary Gynaecology, and Obstetrics, Southern Regional Station of ICAR- National Dairy Research Institute, Bengaluru – 560030 Karnataka
India
Login to access the Email id

Source of Support: The present study was carried out under the project “Molecular markers for improving reproduction in cattle and buffaloes” under the funding of Bill and Melinda Gates Foundation, USA and Indian Council of Agricultural Research - National Dairy Research Institute , Conflict of Interest: None


DOI: 10.4103/2305-0500.294667

Rights and Permissions

Objective: To assess the stability of 10 candidate internal control genes (ICGs), namely GAPDH, ACTB, RPL23, RPS15A, ATPSF1, GLUT5, HMBS, ATP2B4, PPIA, and BRP to normalize the transcriptional data from testes samples of Zebu and crossbred bulls. Methods: Total RNA was isolated from testicular tissue of Zebu and crossbred bulls (n=6 each) between 2-8 years of age. cDNA was synthesized, and the quantitative real-time polymerase chain reaction (PCR) was performed. The cycle threshold values were used for the analysis of the stability of ICGs. Four different statistical algorithms: geNorm, Normfinder, BestKeeper, and RefFinder, were used to assess the stability of these genes. Results: ATPSF1, HMBS, PPIA, and RPS15A were the most reliable and stable ICGs for Zebu testes, and ATPSF1, RPL23, and PPIA for crossbred testes. Conclusions: A panel of stable ICGs (ATPSF1, HMBS, PPIA, RPS15A for Zebu and ATPSF1, RPL23, and PPIA for crossbred) for normalization of gene expression data in testes samples can be helpful for researchers to conduct functional genomics studies at the testicular level in cattle bulls.


[FULL TEXT] [PDF]*
Print this article     Email this article
 Next article
 Previous article
 Table of Contents

 Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
 Citation Manager
 Access Statistics
 Reader Comments
 Email Alert *
 Add to My List *
 * Requires registration (Free)
 

 Article Access Statistics
    Viewed1980    
    Printed73    
    Emailed0    
    PDF Downloaded185    
    Comments [Add]    
    Cited by others 1    

Recommend this journal