Isolation of high spectral quality RNA using run-on gene transcription; Application to gene expression profiling of human brain
Isolation of high spectral quality RNA using run-on gene transcription; Application to gene expression profiling of human brain
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DOI：
10.1007/s10571-005-4035-x
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发表时间：
2005-06-01
影响因子：
4
通讯作者：
Lukiw, WJ
中科院分区：
文献类型：
--
作者：
Cui, JG;Zhao, YH;Lukiw, WJ
1.健康和疾病人脑组织的基因表达研究需要使用完整的、高光谱质量的信使RNA(信使RNA)。短的死后组织，如果控制得当，是大脑mRNA2的重要来源。典型的信使核糖核酸提取方法依赖于从整个细胞或组织中提取总RNA。在这里，我们报告了另一种方法，可以从相同的组织中分离出细胞核，并对其进行编程，以进行连续的基因转录，以产生丰富的mRNA组分。这项新技术比以前的方法有了很大的改进，提供了高质量、高产量的mRNA样本，适合于下游应用，包括使用DNA阵列技术的克隆、测序和基因表达研究。
1. Gene expression studies of human brain tissues in both health and disease require the use of intact, high spectral quality messenger RNA (mRNA). Short post-mortem tissues, when adequately controlled, provide a valuable primary source of brain mRNA.2. Typical mRNA isolation methodologies rely on total RNA extraction from whole cells or tissues. Here we report that as an alternative approach, nuclei can be isolated from the same tissues and programmed for run-on gene transcription to generate enriched mRNA fractions.3. This novel technique represents a significant improvement over previous methods and provides high quality, high yield mRNA samples suitable for downstream applications that include cloning, sequencing and gene expression studies using DNA array technologies.