| 학회 | 한국화학공학회 |
| 학술대회 | 2011년 봄 (04/27 ~ 04/29, 창원컨벤션센터) |
| 권호 | 17권 1호, p.423 |
| 발표분야 | 생물화공 |
| 제목 | Precise multiplex gene expression analysis based on MLPA-CE-SSCP |
| 초록 | Quantification of mRNA provides information crucial for various biological studies. Real-time PCR is known to be the most accurate method for quantifying mRNA, and thus represents the state-of-the-art for gene expression analysis. However, the use of real-time PCR for mRNA quantification is limited to a single target per analytical run because of reductions in quantification power and limitations of fluorescence dyes associated with multiplex applications. Capillary electrophoresis-based single-strand conformation polymorphism analysis (CE-SSCP is an alternative multiplex analysis method. However, CE-SSCP has not been widely used for multiplex applications due to low resolution problem. In this study, we developed high-resolution CE-SSCP system using PEO-PPO-PEO triblock copolymer solution. Moreover, for the multiplex amplification of RNA, modified multiplex ligation-dependent probe amplification (MLPA) was combined with CE-SSCP analysis so that the amount of mRNA could be quantified precisely. We have demonstrated that MLPA-CE-SSCP could be used to monitor expression of 31 metabolic genes of Escherichia coli and 16 genes of Caenorhabditis elegans. |
| 저자 | 신기원1, 황희성2, 정보람1, 정규열2 |
| 소속 | 1시스템생명공학부, 2포항공과대 |
| 키워드 | Expression analysis; Multiplex analysis; MLPA; High-resolution CE-SSCP |
| 원문파일 | 초록 보기 |
