Faculty Collaboration Database

Design of an automated multicapillary instrument with fraction collection for DNA mutation discovery by constant denaturant capillary electrophoresis (CDCE). J Sep Sci 2005 Aug;28(12):1375-89

Date

09/06/2005

Pubmed ID

16138690

DOI

10.1002/jssc.200500023

Scopus ID

2-s2.0-23944499103 (requires institutional sign-in at Scopus site)   12 Citations

Abstract

A fundamental goal ingenomics is the discovery of genetic variation that contributes to disease states or to differential drug responses. Single nucleotide polymorphism (SNP) detection has been the focus of much attention in the study of genetic variation over the last decade. These SNPs typically occur at a frequency greater than 1% in the human genome. Recently, low-frequency alleles are also being increasingly recognized as critical to obtain an improved understanding of the correlation between genetic variation and disease. Although many methods have been reported for the discovery and scoringof SNPs, sensitive, automated, and cost-effective methods and platforms for the discovery of low-frequency alleles are not yet readily available. We describe here an automated multicapillary instrument for high-throughput detection of low-frequency alleles from pooled samples using constant denaturant capillary electrophoresis. The instrument features high optical sensitivity (1 x 10(-12) M fluorescein detection limit), precise and stable temperature control (+/- 0.01degrees C), and automation for sample delivery, injection, matrix replacement, and fraction collection. The capillary array is divided into six groups of four capillaries, each of which can be independently set at any temperature ranging from room temperature to 90 degrees C. The key performance characteristics of the instrument are reported.

Author List

Li Q, Deka C, Glassner BJ, Arnold K, Li-Sucholeiki XC, Tomita-Mitchell A, Thilly WG, Karger BL

Author

Aoy Tomita Mitchell PhD Professor in the Surgery department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Antigens, CD
Antigens, Differentiation
Base Sequence
CTLA-4 Antigen
DNA
Electrophoresis, Capillary
Equipment Design
Genomics
Humans
Methyltransferases
Mutation
Nucleic Acid Denaturation
Optics and Photonics
Polymerase Chain Reaction
Polymorphism, Single Nucleotide
Sensitivity and Specificity
Spectrometry, Fluorescence
Temperature