Annealing control primer system for identification of differentially expressed genes on agarose gels
Annealing control primer system for identification of differentially expressed genes on agarose gels
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DOI：
10.2144/04363st02
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发表时间：
2004-03-01
期刊：
影响因子：
2.7
通讯作者：
Chun, JY
中科院分区：
文献类型：
--
作者：
Kim, YJ;Kwak, CI;Chun, JY
我们开发了GeneFishing技术，这是一种改进的方法，用于使用我们新的退火控制引物（ACP）系统鉴定差异表达基因（DEG）。由于在使用ACP系统的PCR过程中具有高退火特异性，因此将ACP应用于DEG发现产生了可在琼脂糖凝胶上检测到的可再现的、真实的和长的（100 hp至2 kb）PCR产物。为了证明这种基因表达谱分析方法，使用GeneFishing技术，使用从妊娠4.5-18.5天的小鼠孕体组织中分离的总RNA，检测发育期间差异表达的基因。分离得到10个DEG（DEG 1 -10），经北方印迹杂交证实。序列分析表明DEG 6和DEG 10为未知基因。
We developed GeneFishing technology, an improved method for the identification of differentially expressed genes (DEGs) using our novel annealing control primer (ACP) system. Because of high annealing specificity during PCR using the ACP system, the application of the ACP to DEG discovery generates reproducible, authentic, and long (100 hp to 2 kb) PCR products that are detectable on agarose gels. To demonstrate this method for gene expression profiling, GeneFishing technology was used to detect genes that are differentially expressed during development using total RNAs isolated from mouse conceptus tissues at 4.5-18.5 days of gestation. Ten DEGs (DEG1-10) were isolated and confirmed by Northern blot hybridization. The sequence analysis of these DEGs showed that DEG6 and DEG10 are unknown genes.