Unknown

Dataset Information

0

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry.


ABSTRACT: Complex diseases are often underpinned by multiple common genetic variants that contribute to disease susceptibility. Here, we describe a cost-effective tag single nucleotide polymorphism (SNP) approach using a multiplexed genotyping assay with mass spectrometry, to investigate gene pathway associations in clinical cohorts. We investigate the food allergy candidate locus Interleukin13 (IL13) as an example. This method efficiently maximizes the coverage by taking advantage of shared linkage disequilibrium (LD) within a region. Selected LD SNPs are then designed into a multiplexed assay, enabling up to 40 different SNPs to be analyzed simultaneously, boosting cost-effectiveness. Polymerase chain reaction (PCR) is used to amplify the target loci, followed by single nucleotide extension, and the amplicons are then measured using matrix-assisted laser desorption/ionization-time of flight(MALDI-TOF) mass spectrometry. The raw output is analyzed with the genotype calling software, using stringent quality control definitions and cut-offs, and high probability genotypes are determined and output for data analysis.

SUBMITTER: Ashley SE 

PROVIDER: S-EPMC6101967 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry.

Ashley Sarah E SE   Meyer Braydon A BA   Ellis Justine A JA   Martino David J DJ  

Journal of visualized experiments : JoVE 20180621 136


Complex diseases are often underpinned by multiple common genetic variants that contribute to disease susceptibility. Here, we describe a cost-effective tag single nucleotide polymorphism (SNP) approach using a multiplexed genotyping assay with mass spectrometry, to investigate gene pathway associations in clinical cohorts. We investigate the food allergy candidate locus Interleukin13 (IL13) as an example. This method efficiently maximizes the coverage by taking advantage of shared linkage diseq  ...[more]

Similar Datasets

| S-EPMC8632241 | biostudies-literature
| S-EPMC9280723 | biostudies-literature
| S-EPMC7132638 | biostudies-literature
| S-EPMC5241079 | biostudies-literature
| S-EPMC5629267 | biostudies-literature
| S-EPMC2930185 | biostudies-literature
| S-EPMC7710141 | biostudies-literature
| S-EPMC5411034 | biostudies-literature
2005-09-20 | GSE2744 | GEO
| S-EPMC5731921 | biostudies-literature