Annealing control primer system for identification of differentially expressed genes on agarose gels.
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 bp to 2 kb) PCR products that are detectable on agarose gels. To demonstrate this method for gene expression profiling, Gene-Fishing 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.
Kim
Yun-Jee
YJ
Seegene Life Science Laboratory, Seoul, South Korea.
Kwak
Chae-Il
CI
Gu
Young-Yun
YY
Hwang
In-Taek
IT
Chun
Jong-Yoon
JY
D000818:Animals; D002847:Chromatography, Agarose / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D005314:Embryonic and Fetal Development / Q000235:genetics; D005326:Fetal Proteins / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics*; D051379:Mice
D017931:DNA Primers; D005326:Fetal Proteins