competitive rt-pcr strategy for quantitative evaluation of the expression of tilapia (oreochromis niloticus) growth hormone receptor type i竞争性rt - pcr定量评价策略表达式的罗非鱼(oreochromis niloticus)生长激素受体i型.pdfVIP

Competitive RT-PCR Strategy for Quantitative Evaluation of the Expression of Tilapia (Oreochromis niloticus) Growth Hormone Receptor Type I Alina Rodríguez-Mallon, Yamilet Cárdenas, Juana María Lugo, Aymé Oliva, Antonio Morales, and Mario Pablo Estrada Abstract Quantization of gene expression requires that an accurate measurement of a specific transcript is made. In this paper, a quantitative reverse transcription-polymerase chain reaction (RT-PCR) by competition for tilapia growth hormone receptor type I is designed and validated. This experimental procedure was used to determine the abundance of growth hormone receptor type I transcript in different tilapia tissues. The results obtained with this developed competitive RT-PCR were similar to real-time PCR results reported recently. This proto- col provides a reliable alternative, but less expensive than real-time PCR to quantify specific genes. Keywords: Tilapia, Receptors, Somatostatin. Abbreviations Ab Antibiotic C Competitor DNA Deoxyribonucleic acid GH Growth hormone GHR Growth hormone receptor mRNA Messenger ribonucleic acid o.n. Overnight PCR Polymerase chain reaction RNA Ribonucleic acid RT Reverse transcription T Target tiGHR I Tilapia growth hormone receptor type I T Melting temperature m Electronic supplementary material: The online version of this article (DOI:10.1007/s12575-009-9002-3) contains supplementary material, which is available to author