title: Variant Calling From Next Generation Sequence Data.
abstract: The use of next generation nucleotide sequencing to discover and genotype small sequence variants has led to numerous insights into the molecular causes of various diseases. This chapter describes the use of freely available software to align next generation sequencing reads to a reference and then to use the resulting alignments to call, annotate, view, and filter small sequence variants. The suggested variant calling workflow includes read alignment with novoalign, the removal of polymerase chain reaction duplicate sequences with samtools or bamUtils, and the detection of variants with Freebayes or bam2mpg software. ANNOVAR is then used to annotate the predicted variants using gene models, population frequencies, and predicted mutation severity, producing variant files which can be viewed and filtered with the variant display tool VarSifter.
authors.affiliation: National Human Genome Research Institute, Rockville, MD, USA.
authors.forename: Nancy F
authors.initials: NF
authors.lastname: Hansen
mesh_terms: D019295:Computational Biology / Q000379:methods*; D030541:Databases, Genetic; D059014:High-Throughput Nucleotide Sequencing; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods*; D012984:Software; D064878:Web Browser
keywords: Annotation; Genotyping; Next generation sequencing; Variant detection; Whole exome