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上海玉博生物科技有限公司
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文献和实验using the AllPrep DNA/RNA 96 Kit. Real-time RT-PCR analysis of the PGK1 transcript was performed using a primer-probe set that could amplify and detect both mRNA and genomic DNA sequences. The flat amplification plot for the no RT control (-RT) indicates
by reverse transcriptase (RT)-PCR from highly specialized cells (e.g. isolated from a heterogenous sample by immunomagnetic separation). In addition, the protocol has been successfully used to isolate mRNA from a wide variety of tissues of mammalian, fish
(A)+RNA。不管起始模板是总RNA还是poly(A)+ RNA,都可以检测到扩增结果(图2)。另外,分离poly(A)+ RNA会导致样品间mRNA丰度的波动变化,从而使信息的检出和定量产生偏差。然而,当分析稀有mRNA时,poly(A)+ RNA会增加检测的灵敏度。 图2. 总RNA和poly(A)+ RNA在RT-PCR中的比较 以5或1μg Hela细胞总RNA(分别为泳道1和2)或500ng和50ng Hela细胞poly(A)+ RNA(分别为泳道
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