Ishimaru T, Hirose T, Matsuda T, Goto A, Takahashi K, Sasaki H, Terao T, Ishii R, Ohsugi R, Yamagishi T. Expression profiling of genes involved in starch synthesis in sink and source organs of rice. Ohda T, Francisco PB, Sawada T, Hirose T, Terao T, Satoh H, Nakamura Y. Quantification of mRNA using real-time RT-PCR. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems.
Basic principles of real-time quantitative PCR. Genome Res.1996 6:986-94.Īrya M, Shergill IS, Williamson M, Gommersall L, Arya N, Patel HR. Heid CA, Stevens J, Livak KJ, Williams PM. Real-time quantitative RT-PCR after laser assisted cell picking. Therefore, there is a dire need to validate a set of housekeeping genes in the species under study and in various tissues depending on the experimental design to obtain valid and reproducible results.įink L, Seeger W, Ermert L, Hanze J, Stahl U, Grimminger F, Kummer K, Bohle RM. Using the geometric mean of the three most stable genes as a housekeeping gene can also solve this problem to some extent. Moreover, it is recommended to use more than one housekeeping gene for relative quantification.
To solve this problem, various types of user-friendly software are now available for validation of housekeeping genes. Use of housekeeping genes without proper validation may lead to erroneous results and misinterpretation of data. In light of the literature reviewed here, it is clear that expression of most of the genes varied in different physiological conditions, under different experimental conditions, and in different gender.
Expression of housekeeping genes were thought to be constant in different tissues under various physiological and experimental conditions. Relative quantification of mRNA through real-time qPCR requires suitable housekeeping genes/reference genes. Its use in aquaculture and study of fish physiology is increasing day by day. Real-time polymerase chain reaction (qPCR) is a reliable tool for gene expression analysis in various organisms.
0 kommentar(er)
