RT-PCR测定mRNA的荧光定量分析

来源 :生物化学与生物物理学报 | 被引量 : 0次 | 上传用户:herry
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利用反转录毛细管PCR技术,可合成代表特异mRNA的双链DNA。在一定循环数内,PCR产物的量与其模板cDNA即反转录中相应mRNA的浓度有关。溴乙锭嵌入DNA双螺旋后,其荧光量子产率大为增加,因此可通过利用处在适当PCR循环数中的cDNA浓度与在优选的激发光谱与发射波长下其荧光强度的线性关系,计算出所检测的mRNA表达量。 Using reverse-transcription capillary PCR, double-stranded DNA representing specific mRNAs can be synthesized. Within a certain number of cycles, the amount of PCR product is related to the concentration of the corresponding mRNA in the reverse transcription of its template cDNA. When ethidium bromide is inserted into a DNA duplex, its fluorescence quantum yield is greatly increased and can therefore be calculated by using the linear relationship between the concentration of cDNA in the appropriate PCR cycles and its fluorescence intensity at the preferred excitation spectrum versus emission wavelength The detected amount of mRNA expression was measured.
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