Isolation of microarray-quality RNA from primary human cells after intracellular immunostaining and fluorescence-activated cell sorting.
Isolation of microarray-quality RNA from primary human cells after intracellular immunostaining and fluorescence-activated cell sorting.
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DOI：
10.1016/j.jim.2013.02.003
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发表时间：
2013-05-31
影响因子：
2.2
通讯作者：
Romerio, Fabio
中科院分区：
文献类型：
--
作者：
Iglesias-Ussel, Maria;Marchionni, Luigi;Romerio, Fabio
微阵列使从极其广泛的来源进行高通量、全基因组的RNA表达分析成为可能。这项技术依赖于为这类应用获得足够数量和质量的RNA的能力。虽然有一些方法可以绕过前者的限制，但在处理一些生物样品时，适合于微阵列分析的RNA的回收仍然是一个主要问题，特别是那些用核酸修饰有机试剂处理和保存的样品。在本报告中，我们描述了一种从荧光激活细胞分选纯化的细胞中提取适合于微阵列分析的RNA的方法，所述细胞经过固定、渗透和用荧光素偶联抗体进行细胞内染色。我们表明--尽管从这些样品中分离的RNA出现了一些降解--但它在微阵列分析中表现得非常好。我们在这里描述的方法使其可用于全基因组表达谱分析，迄今仅限于单基因分析的各种生物样本。
Microarrays have made it possible to perform high-throughput, genome-wide analyses of RNA expression from an extremely wide range of sources. This technology relies on the ability to obtain RNA of sufficient quantity and quality for this type of application. While there are means to circumvent limitations in the former, recovery of RNA suitable for microarray analysis still represents a major issue when working with some biological samples, particularly those treated with and preserved in nucleic acid-modifying organic reagents. In the present report we describe a procedure for the isolation of RNA suitable for microarray analysis from cells purified by fluorescence-activated cell sorting after fixation, permeabilization and intracellular staining with fluorochrome-conjugated antibodies. We show that – although the RNA isolated from these samples presented some degradation – it performed remarkably well in microarray analysis. The method we describe here makes it available to genome-wide expression profiling a variety of biological samples that so far were confined to single-gene analysis.
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影响因子：
56.9
作者：
Carninci, P;Kasukawa, T;Hayashizaki, Y
通讯作者：
Hayashizaki, Y
影响因子：
--
作者：
Schroeder A;Mueller O;Stocker S;Salowsky R;Leiber M;Gassmann M;Lightfoot S;Menzel W;Granzow M;Ragg T
通讯作者：
Ragg T
DOI：
10.1158/1078-0432.ccr-09-1631
发表时间：
2009-11-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Conway C;Mitra A;Jewell R;Randerson-Moor J;Lobo S;Nsengimana J;Edward S;Sanders DS;Cook M;Powell B;Boon A;Elliott F;de Kort F;Knowles MA;Bishop DT;Newton-Bishop J
通讯作者：
Newton-Bishop J
影响因子：
11.2
作者：
Li, HR;Wang-Rodriguez, J;Fan, JB
通讯作者：
Fan, JB
影响因子：
56.9
作者：
Bertone, P;Stolc, V;Snyder, M
通讯作者：
Snyder, M