Semiquantitative RT-PCR analysis to assess the expressionlevels of iNOS in chicken macrophages

Inos is inducible by a variety of factors related to inflammation and referred to asinducible nos(inos). it is regulated at the level of gene expression; once expressed, it producesno at a high rate. inos gene-expression profiling is an important tool in understandingmolecular markers of the responses of cells and tissues to external factors. in this article asemiquantitative reverse transcription-polymerase chain reaction (rt-pcr) protocol wasoptimized to extract rna (ribonucleic acid) from chicken spleen and to measure the expressionlevels of inos mrnas from each sample. detailed procedure was described for the analysis ofinos levels. β-actin was used as an internal control to normalize for sample to sample variationsin total rna amounts and for reaction efficiency. co-amplification of the inos gene withhousekeeping gene (β-actin) provides a quantitative result. changes in gene expression levelmay be monitored, while avoiding sample-to-sample loading variation.