High Resolution Analysis of Point Mutations by Constant Denaturant Capillary Electrophoresis (CDCE)
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In recent years, the need for techniques capable of detecting and identifying point mutants has increased dramatically in
the fields of genomics, cancer research, and molecular diagnostics. The large arsenal of methods for mutation detection ranges
from direct sequencing and array hybridization to allele-specific polymerase chain reaction (PCR). CDCE (1) holds a unique position within this list of techniques due to its high flexibility. Having set up a CDCE system, one can
use it to solve a wide range of tasks. At one extreme, CDCE is capable of detecting mutants at very low frequencies (as low
as 10-6) (2,3), in which it rivals the high sensitivity mutation detection methods like allele specific PCR. At the other extreme, CDCE
is capable of identifying mutants with precision comparable to that of direct sequencing (4). In addition, the instrument for CDCE does not need to be dedicated to one purpose. In fact, most capillary DNA electrophoresis
systems, such as DNA sequencing capillary instruments, can be adapted for CDCE separations.
Affiliation(s): (5) Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA
(6) Fred Hutchinson Cancer Research Center, Seattle, WA
(7) Division of Bioengineering and Environmental Health, Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA
(8) Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA
(6) Fred Hutchinson Cancer Research Center, Seattle, WA
(7) Division of Bioengineering and Environmental Health, Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA
(8) Center for Environmental Health Sciences, Massachusetts Institute of Technology, Cambridge, MA
Book Title: Capillary Electrophoresis of Nucleic Acids: Volume II: Practical Applications of Capillary Electrophoresis
Series: Methods in Molecular Biology | Volume: 163 | Pub. Date: Jan-19-2001 | Page Range: 57-72 | DOI: 10.1385/1-59259-116-7:57
Subject: Biochemistry
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