Expression stability of two housekeeping genes (18S rRNA and G3PDH) during in vitro maturation of follicular oocytes in buffalo (Bubalus bubalis)

Ajay Pal Singh Aswal; Sarvesh Raghav; Sachinandan De; Manish Thakur; Surender Lal Goswami; Tirtha Kumar Datta

doi:10.1016/j.anireprosci.2007.04.012

Expression stability of two housekeeping genes (18S rRNA and G3PDH) during in vitro maturation of follicular oocytes in buffalo (Bubalus bubalis)

Anim Reprod Sci. 2008 Jan 15;103(1-2):164-71. doi: 10.1016/j.anireprosci.2007.04.012. Epub 2007 Apr 22.

Authors

Ajay Pal Singh Aswal¹, Sarvesh Raghav, Sachinandan De, Manish Thakur, Surender Lal Goswami, Tirtha Kumar Datta

Affiliation

¹ Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India.

PMID: 17537596
DOI: 10.1016/j.anireprosci.2007.04.012

Abstract

The present study was undertaken to evaluate the expression stability of two housekeeping genes (HKGs), 18S rRNA and G3PDH during in vitro maturation (IVM) of oocytes in buffalo, which qualifies their use as internal controls for valid qRT-PCR estimation of other oocyte transcripts. A semi quantitative RT-PCR system was used with optimised qRT-PCR parameters at exponential PCR cycle for evaluation of temporal expression pattern of these genes over 24 h of IVM. 18S rRNA was found more stable in its expression pattern than G3PDH.

MeSH terms

Animals
Buffaloes / genetics
Buffaloes / metabolism
Buffaloes / physiology*
Female
Gene Expression
Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / biosynthesis*
Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / genetics
Oocytes / enzymology
Oocytes / physiology*
Ovarian Follicle / enzymology
Ovarian Follicle / physiology*
RNA, Messenger / biosynthesis
RNA, Messenger / genetics
RNA, Ribosomal, 18S / biosynthesis*
RNA, Ribosomal, 18S / genetics
Regression Analysis
Reverse Transcriptase Polymerase Chain Reaction / methods

Substances

RNA, Messenger
RNA, Ribosomal, 18S
Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)