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首页> 外文期刊>Brain research. Brain research protocols >Detection and semi-quantitative determination of low abundance GFAP mRNA in mouse brain by capillary electrophoresis coupled with laser-induced fluorescence.
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Detection and semi-quantitative determination of low abundance GFAP mRNA in mouse brain by capillary electrophoresis coupled with laser-induced fluorescence.

机译:毛细管电泳结合激光诱导的荧光检测和半定量测定小鼠脑中低丰度GFAP mRNA。

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摘要

The sensitivity of capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) was compared with conventional agarose gel electrophoresis-ethidium bromide-UV method (AE-EUV) for detection and semi-quantitative determination of GFAP mRNA in mouse brain. GFAP expression was induced by the neurotoxin MPTP in C57BL mice. Serially diluted RNA samples (0.0003, 0.003, 0.03, 0.3, and 3 mug total RNA) were subjected to RT-PCR and analyzed by both procedures. The integrated pixel density (AE-EUV) and peak area (CE-LIF) were directly proportional to the amount of RNA. However, the observed high sensitivity of CE-LIF suggests its potential application for detection and semi-quantitative determination of low-abundance mRNA transcripts.
机译:比较了毛细管电泳与激光诱导荧光(CE-LIF)的灵敏度与常规琼脂糖凝胶电泳-溴化乙锭-UV方法(AE-EUV)在小鼠脑中GFAP mRNA的检测和半定量测定。在C57BL小鼠中神经毒素MPTP诱导GFAP表达。对连续稀释的RNA样品(0.0003、0.003、0.03、0.3和3杯总RNA)进行RT-PCR,并通过两种方法进行分析。积分像素密度(AE-EUV)和峰面积(CE-LIF)与RNA量成正比。然而,观察到的CE-LIF的高灵敏度表明它在检测和半定量测定低丰度mRNA转录本方面的潜在应用。

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