METHOD FOR IDENTIFYING INTRACELLULAR EXPRESSION LEVEL OF MRNAS OF E6 AND E7 PROTEINS USING POLYMERASE CHAIN REACTION AND SEQUENCE SPECIFIC PROBE

摘要

PURPOSE: A method for identifying intracellular expression level of mRNAs of E6 and E7 proteins is provided which is time and cost effective by using PCR and a sequence specific probe without contamination of a PCR product. CONSTITUTION: A method for identifying intracellular expression level of mRNA comprises the steps of: designing a primer which is specifically bound to mRNA of E6 and E7 proteins and binding it to the proteins; performing PCR for mRNA, which is isolated from sample, using the primer; designing a sequence specific probe which is specifically bound to the PCR product followed by binding biotin as a marker to the probe; binding the probe to microwell plate which is coated with a streptavidin; heating the PCR product to denature and then hybridizing in the microwell plate; adding an anti-digoxigenin antibody attached an enzyme and a color reagent to the microwell plate; and analyzing the coloring using a spectrophotometer.