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Method for quantitative measuring short RNA using amplified DNA fragment length polymorphism
Method for quantitative measuring short RNA using amplified DNA fragment length polymorphism
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机译:利用扩增的DNA片段长度多态性定量测定短RNA的方法
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摘要
The present invention relates to the technical field of molecular biology, provides a method for measuring short RNA using amplified DNA fragment length polymorphism, and comprises the following steps: first using at least two synthesized miRNAs as the internal measurement standard, said synthesized miRNAs containing no natural homologous sequence in comparison with the short RNA to be measured, and mixing the synthesized miRNAs using different molecule numbers so as to form a dynamic miRNA standard molecular gradient; mixing the same quantity of the dynamic miRNA standard with the short RNA to be measured, and performing RNA reverse transcription, cDNA tailing, PCR synchronous amplification, and fluorescent quantitative analysis on the length polymorphism fragment of the PCR product DNA so as to measure the relative ratio of the fluorescence intensity of the DNA fragment produced by the amplification of the short RNA to be measured to the dynamic miRNA standard fluorescence intensity gradient.
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