Application of card blot as a semi-quantitative detection genes expression

The purpose of this research is to invent a break-trough methods of genes expression which are based on RNA-DNA hybridization, through a sensitive, efficient and cost effective way. In this research, biotin labeled probe were used to measure bc1-2 and c-myc niRNA genes expressions of peripheral blood lymphocyte from patients of Infectious Mononucleosis (IM), Nasopharyngeal Carcinoma (NPC) and Non-diagnostic asymptomatic (NDA). The blotting process was brought about to attach the total RNA sample at the nitrocellulose membrane on card blot through vacuum process, continued by washing and baking at 80 ° C degrees for a period of 1 hour. Hybridization was undergone using bc1-2 and c-myc genes DNA fragment biotin labeled probe. Indirect detection system utilizing Streptavidin-Avidin - Horse Raddish Peroxidase (SA-HRP) to bind the biotin, was amplified with Biotynil-Tyramide (BT) and Diaminobenzidine (DAB) chromogenic substrate that exists in Tyramide Signal AmplificationTM-Indirect (ISH). Result of this research showed that card blot can be used in detecting gene on the RNA level with a sum of 20 ng/sample rapidly and with an effective cost. Keywords: card blot dot blot â TS