Chemiluminescent analysis of gene expression on high-density filter arrays
Chemiluminescent analysis of gene expression on high-density filter arrays
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DOI：
10.1177/002215549904700307
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发表时间：
1999-03-01
影响因子：
3.2
通讯作者：
Vernon, SD
中科院分区：
文献类型：
--
作者：
Rajeevan, MS;Dimulescu, IM;Vernon, SD
我们已经优化了使用高密度滤光片阵列（HDFAs）的荧光分析基因表达的条件。通过优化cDNA探针合成、杂交和检测参数，实现了高灵敏度和特异性。荧光表达谱反映了从不同来源（胎盘和角质形成细胞）分离的转录本的预期差异。我们估计低丰度信息的检测限为每个细胞1-15个转录本，灵敏度与微阵列格式的报告相当，超过放射自显影HDFA的报告。该方法允许短的暴露时间和探针的重复使用。它同样适用于cDNA文库的差异筛选和差异显示PCR等技术。
We have optimized conditions for the chemiluminescent analysis of gene expression using high-density filter arrays (HDFAs). High sensitivity and specificity were achieved by optimizing cDNA probe synthesis, hybridization, and detection parameters. The chemiluminescent expression profile reflected expected differences in the transcripts isolated from different sources (placenta and keratinocytes). We estimated the detection limit for low-abundance message to be 1-15 transcripts per cell, a sensitivity rivaling that reported for microarray formats and exceeding that reported for autoradiographic HDFAs. The method allows for short exposure times and reuse of probe. It should be equally applicable to techniques such as differential screening of cDNA libraries and differential display PCR.