Method for Quantitative Measuring Short RNA Using Amplified DNA Fragment Length Polymorphism

A fluorescent capillary electrophoresis method of quantitatively measurement of short-chain RNAs using amplified fragment length polymorphism of DNA, characterized in that:

• said method comprises the following steps of:
• first, utilizing at least two types of synthetic microRNA which has no natural homologous sequence when compared to a short-chain RNA for testing as an internal standard for measurement;
• and mixing the synthetic microRNA, which is used as the internal standard, with different molecule numbers to form a standard molecular gradient of dynamic microRNA;
• then, mixing the short-chain RNA for testing with an equal amount of the dynamic microRNA standard;
• processing RNA reverse transcription, cDNA tailing, PCR synchronous amplification and fluorescence quantitative analysis of DNA length polymorphism fragment of PCR products to measure a relative ratio of fluorescence intensity of DNA fragment produced from the amplification of the short-chain RNA for testing based on the standard fluorescence intensity gradient of dynamic microRNA, thereby achieving a relative quantification of the short-chain RNA for testing,
• utilizing the short-chain RNA for testing as a template to synthesis short-chained RNA reference;
• then determining a relative ratio of RNA reference for testing at different molecular numbers on the standard fluorescence intensity gradient of dynamic microRNA, thereby obtaining a calibration curve of number of molecules—relative intensity of the RNA reference for testing and calculating an absolute number of molecules of the short-chain RNA for testing in the test sample by utilizing the ratio of relative intensity of the short-chain RNA through the calibration curve,
• wherein the short-chain RNA for testing refers to microRNA (miRNA).