Single-stranded conformational polymorphism analysis using automated capillary array electrophoresis apparatuses

被引:24
作者
Baba, S
Kukita, Y
Higasa, K
Tahira, T
Hayashi, K
机构
[1] Kyushu Univ, Div Genome Anal, Res Ctr Genet Informat, Med Inst Bioregulat, Fukuoka 812, Japan
[2] Univ Tokyo, Tokyo, Japan
关键词
D O I
10.2144/03344st03
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We describe a new environment of a single-stranded conformational polymorphism (SSCP) analysis using automated capillary array sequencers (e.g., ABI PRISM(R) 3100 and 3700). In this environment, electrophoretic conditions, settings for instrument management, and software for data analysis are adjusted for SSCP analysis. Highly reproducible results are obtained with this new system, and fragments with mutations and/or polymorphisms in different capillaries or different runs can be reliably detected. The relative peak heights between alleles are quantitative and reproducible between runs, and so allele frequencies of single nucleotide polymorphisms can be accurately estimated by a pooled DNA strategy. The method allows unattended, low-cost, and quantitative SSCP analysis using instruments that are widely accessible.
引用
收藏
页码:746 / 750
页数:5
相关论文
共 20 条
[1]  
Brooks Lisa D, 2003, Methods Mol Biol, V212, P1
[2]   Base-calling of automated sequencer traces using phred.: II.: Error probabilities [J].
Ewing, B ;
Green, P .
GENOME RESEARCH, 1998, 8 (03) :186-194
[3]   Base-calling of automated sequencer traces using phred.: I.: Accuracy assessment [J].
Ewing, B ;
Hillier, L ;
Wendl, MC ;
Green, P .
GENOME RESEARCH, 1998, 8 (03) :175-185
[4]   Consed: A graphical tool for sequence finishing [J].
Gordon, D ;
Abajian, C ;
Green, P .
GENOME RESEARCH, 1998, 8 (03) :195-202
[5]   USE OF LABELED PRIMERS IN POLYMERASE CHAIN-REACTION (LP-PCR) FOR A RAPID DETECTION OF THE PRODUCT [J].
HAYASHI, K ;
ORITA, M ;
SUZUKI, Y ;
SEKIYA, T .
NUCLEIC ACIDS RESEARCH, 1989, 17 (09) :3605-3605
[6]  
Hayashi K, 1991, PCR Methods Appl, V1, P34
[7]   Software for machine-independent quantitative interpretation of SSCP in capillary array electrophoresis (QUISCA) [J].
Higasa, K ;
Kukita, Y ;
Baba, S ;
Hayashi, K .
BIOTECHNIQUES, 2002, 33 (06) :1342-1348
[8]   PRODUCTION OF SINGLE-STRANDED-DNA TEMPLATES BY EXONUCLEASE DIGESTION FOLLOWING THE POLYMERASE CHAIN-REACTION [J].
HIGUCHI, RG ;
OCHMAN, H .
NUCLEIC ACIDS RESEARCH, 1989, 17 (14) :5865-5865
[9]   One-tube post-PCR fluorescent labeling of DNA fragments [J].
Inazuka, M ;
Tahira, T ;
Hayashi, K .
GENOME RESEARCH, 1996, 6 (06) :551-557
[10]   A streamlined mutation detection system: Multicolor post-PCR fluorescence labeling and single-strand conformational polymorphism analysis by capillary electrophoresis [J].
Inazuka, M ;
Wenz, HM ;
Sakabe, M ;
Tahira, T ;
Hayashi, K .
GENOME RESEARCH, 1997, 7 (11) :1094-1103