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Integrated platform for detection of DNA sequence variants using capillary array electrophoresis

Journal Article · · Electrophoresis
OSTI ID:931190
 [1];  [1];  [2];  [2]
  1. SpectruMedix, State College, PA
  2. ORNL
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We have developed a highly versatile platform that performs temperature gradient capillary electrophoresis (TGCE) for mutation/single-nucleotide polymorphism (SNP) detection, sequencing and mutation/SNP genotyping for identification of sequence variants on an automated 24-, 96- or 192-capillary array instrument. In the first mode, multiple DNA samples consisting of homoduplexes and heteroduplexes are separated by CE, during which a temperature gradient is applied that covers all possible temperatures of 50% melting equilibrium (Tms) for the samples. The differences in Tms result in separation of homoduplexes from heteroduplexes, thereby identifying the presence of DNA variants. The sequencing mode is then used to determine the exact location of the mutation/SNPs in the DNA variants. The first two modes allow the rapid identification of variants from the screening of a large number of samples. Only the variants need to be sequenced. The third mode utilizes multiplexed single-base extensions (SBEs) to survey mutations and SNPs at the known sites of DNA sequence. The TGCE approach combined with sequencing and SBE is fast and cost-effective for high-throughput mutation/SNP detection.
Research Organization:
Oak Ridge National Laboratory (ORNL); Mouse Genetics Research Facility
Sponsoring Organization:
SC USDOE - Office of Science (SC)
DOE Contract Number:
AC05-00OR22725
OSTI ID:
931190
Journal Information:
Electrophoresis, Journal Name: Electrophoresis Vol. 23
Country of Publication:
United States
Language:
English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Capillary array electrophoresis
DETECTION
DNA
ELECTROPHORESIS
Genotyping
MELTING
MUTATIONS
Mutation detection
Sequencing
TEMPERATURE GRADIENTS