Impact of RNA extraction from limited samples on microarray results.

Impact of RNA extraction from limited samples on microarray results.
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从有限样品中提取 RNA 对微阵列结果的影响。

DOI:
10.2144/03355st04
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发表时间:
2003
期刊:
影响因子:
2.7
通讯作者:
S. Vernon
S. Vernon
中科院分区:
工程技术4区
文献类型:
--
作者:
Hanna Ojaniemi;B. Evengård;Daisy R. Lee;E. Unger;S. Vernon

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为了将微阵列技术引入诊断领域,需要了解并尽量减少样品制备和 RNA 提取等技术参数的影响。我们评估了两种 RNA 提取方法、DNase 处理和杂交 cDNA 探针的量对微阵列结果的影响。两种 RNA 提取方法的结果相当,尽管其中一种方法会产生残留 DNA,对微阵列结果略有影响。只需 1 微克总 RNA 即可用于合成 cDNA 探针,并产生与使用 5 微克总 RNA 产生的基因表达谱相似的基因表达谱,尽管总体信号强度较低。这些实验表明,微阵列技术对于在诊断环境中使用有限的临床样本具有巨大的前景。
To move microarray technology into the diagnostic realm, the impact of technical parameters, such as sample preparation and RNA extraction, needs to be understood and minimized. We evaluated the impact of two RNA extraction methods, DNase treatment and the amount of hybridized cDNA probe, on the outcome of microarray results. The results for both RNA extraction methods were comparable, although one method resulted in residual DNA that slightly affected the microarray results. As little as one microgram of total RNA could be used to synthesize a cDNA probe and resulted in a gene expression profile that was similar to one produced using 5 micrograms total RNA, even though the overall signal intensity was lower. These experiments illustrate that microarray technology holds great promise for the use of limited clinical samples in the diagnostic setting.
DOI: 10.2144/00293bi01
发表时间: 2000-09-01
期刊: BIOTECHNIQUES
影响因子: 2.7
作者:
Hegde, P;Qi, R;Quackenbush, J
通讯作者: Quackenbush, J
从盲鳗粘液腺中制备 Poly(A) mRNA。
DOI: 10.1080/00327488808062513
发表时间: 1988
期刊: Preparative biochemistry
影响因子: --
作者:
Gadbois,DM;Salo,WL;Ann,DK;Downing,SW;Carlson,DM
通讯作者: Carlson,DM