| 학회 | 한국화학공학회 |
| 학술대회 | 2010년 가을 (10/21 ~ 10/22, 대전컨벤션센터) |
| 권호 | 16권 2호, p.1552 |
| 발표분야 | 생물화공 |
| 제목 | Precise multiplex RNA quantification method based on MLPA-CE-SSCP |
| 초록 | Quantification of RNA provides information crucial for various biological studies. Real-time PCR is known to be the most accurate method for quantifying nucleic acids, and thus represents the state-of-the-art for RNA quantification. However, the use of real-time PCR for RNA 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 RNA quantification method. However, CE-SSCP has not been widely used for multiplex RNA quantification 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 initial amount of RNA could be quantified precisely. We have demonstrated that MLPA-CE-SSCP could be used to monitor expression of 38 metabolic genes of Escherichia coli and 20 Arabidopsis response regulators. |
| 저자 | 신기원, 황희성, 정보람, 정규열 |
| 소속 | 포항공과대 |
| 키워드 | mRNA quantification; MLPA; CE-SSCP; multiplex analysis |
| 원문파일 | 초록 보기 |
