Annealing control primer system for identification of differentially expressed genes on agarose gels

被引：150

作者：

Kim, YJ ^{[1
]}

Kwak, CI ^{[1
]}

Gu, YY ^{[1
]}

Hwang, IT ^{[1
]}

Chun, JY ^{[1
]}

机构：

[1] Seegene Life Sci Lab, Seoul 135090, South Korea

来源：

BIOTECHNIQUES | 2004年 / 36卷 / 03期

关键词：

D O I：

10.2144/04363ST02

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：424 / +

页数：6

共 20 条

[11] FLUORESCENT DIFFERENTIAL DISPLAY - ARBITRARILY PRIMED RT-PCR FINGERPRINTING ON AN AUTOMATED DNA SEQUENCER [J].