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The Advantage of Absolute Quantification in Comparative Hormone Research as Indicated by a Newly Established Real‐Time RT‐PCR: GH, IGF‐I, and IGF‐II Gene Expression in the Tilapia, Oreochromis niloticus
Author(s) -
EPPLER ELISABETH,
CAELERS ANTJE,
BERISHVILI GIORGI,
REINECKE M.NFRED
Publication year - 2005
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1327.047
Subject(s) - real time polymerase chain reaction , gene , gene expression , growth hormone , primer (cosmetics) , biology , tilapia , cloning (programming) , messenger rna , microbiology and biotechnology , computational biology , hormone , fish <actinopterygii> , genetics , chemistry , computer science , endocrinology , organic chemistry , fishery , programming language
A bstract : We have developed a real‐time RT‐PCR that absolutely quantifies the gene expression of hormones using the standard curve method. The method avoids cloning procedures by using primer extension to create templates containing a T7 promoter gene sequence. It is rapid since neither separate reverse transcriptions nor postamplification steps are necessary, and its low detection level (2 pg/μg total RNA) allows precise absolute quantification. Using the method, we have quantified the gene expression of GH, IGF‐I, and IGF‐II in the tilapia.