Genetic mutation analysis by CE with LIF detection using inverse-flow derivatization of DNA fragments.
Electrophoresis. 2006 Oct; 27(19):3846-55.E

Abstract

Inverse-flow derivatization is a novel approach to obtain fluorescent DNA derivatives in DNA analysis based on CE with LIF detection. In the present work, we want to explore the feasibility of the application of this method into the mutation detection based on constant denaturant capillary electrophoresis (CDCE) and SSCP analysis. The DNA fragments were first amplified by PCR using a pair of common primers without fluorescent label, and then the mutations were determined by CDCE or SSCP analysis based on CE-LIF with inverse-flow derivatization of DNA fragments. The experimental conditions were investigated systematically, and different labeling modes including inverse-flow derivatization, on-column derivatization and fluorescent labeled primer technique were compared. The inverse-flow derivatization was successfully used in the detection of C677T mutation in the methylenetetrahydrofolate reductase gene by CDCE or SSCP analysis. Our preliminary results demonstrate that inverse-flow derivatization is very simple, inexpensive and sensitive and well suitable for the genetic analysis in clinical diagnosis.

Authors+Show Affiliations

Sang F

College of Chemistry and Chemical Engineering, Shanghai Jiaotong University, Shanghai, P. R. China.

Ren H

No affiliation info available

Ren J

No affiliation info available

MeSH

DNADNA Mutational AnalysisElectrophoresis, CapillaryFluorescenceFluorescent DyesHumansLasersMethylenetetrahydrofolate Reductase (NADPH2)MutationNucleic Acid DenaturationSpectrometry, Fluorescence

Pub Type(s)

Evaluation Study

Journal Article

Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16941450

Citation

Sang, Fuming, et al. "Genetic Mutation Analysis By CE With LIF Detection Using Inverse-flow Derivatization of DNA Fragments." Electrophoresis, vol. 27, no. 19, 2006, pp. 3846-55.

Sang F, Ren H, Ren J. Genetic mutation analysis by CE with LIF detection using inverse-flow derivatization of DNA fragments. Electrophoresis. 2006;27(19):3846-55.

Sang, F., Ren, H., & Ren, J. (2006). Genetic mutation analysis by CE with LIF detection using inverse-flow derivatization of DNA fragments. Electrophoresis, 27(19), 3846-55.

Sang F, Ren H, Ren J. Genetic Mutation Analysis By CE With LIF Detection Using Inverse-flow Derivatization of DNA Fragments. Electrophoresis. 2006;27(19):3846-55. PubMed PMID: 16941450.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Genetic mutation analysis by CE with LIF detection using inverse-flow derivatization of DNA fragments. AU - Sang,Fuming, AU - Ren,Haixia, AU - Ren,Jicun, PY - 2006/8/31/pubmed PY - 2007/1/5/medline PY - 2006/8/31/entrez SP - 3846 EP - 55 JF - Electrophoresis JO - Electrophoresis VL - 27 IS - 19 N2 - Inverse-flow derivatization is a novel approach to obtain fluorescent DNA derivatives in DNA analysis based on CE with LIF detection. In the present work, we want to explore the feasibility of the application of this method into the mutation detection based on constant denaturant capillary electrophoresis (CDCE) and SSCP analysis. The DNA fragments were first amplified by PCR using a pair of common primers without fluorescent label, and then the mutations were determined by CDCE or SSCP analysis based on CE-LIF with inverse-flow derivatization of DNA fragments. The experimental conditions were investigated systematically, and different labeling modes including inverse-flow derivatization, on-column derivatization and fluorescent labeled primer technique were compared. The inverse-flow derivatization was successfully used in the detection of C677T mutation in the methylenetetrahydrofolate reductase gene by CDCE or SSCP analysis. Our preliminary results demonstrate that inverse-flow derivatization is very simple, inexpensive and sensitive and well suitable for the genetic analysis in clinical diagnosis. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/16941450/Genetic_mutation_analysis_by_CE_with_LIF_detection_using_inverse_flow_derivatization_of_DNA_fragments_ L2 - https://doi.org/10.1002/elps.200600160 DB - PRIME DP - Unbound Medicine ER -

Tags

Genetic mutation analysis by CE with LIF detection using inverse-flow derivatization of DNA fragments.Electrophoresis. 2006 Oct; 27(19):3846-55.E