Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.

RNA sequencing (RNAseq) samples the majority of expressed genes infrequently, owing to the large size, complex splicing and wide dynamic range of eukaryotic transcriptomes. This results in sparse sequencing coverage that can hinder robust isoform assembly and quantification. RNA capture sequencing (...

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Detalhes bibliográficos
Principais autores: Mercer, T, Clark, M, Crawford, J, Brunck, M, Gerhardt, D, Taft, R, Nielsen, L, Dinger, M, Mattick, J
Formato: Journal article
Idioma:English
Publicado em: Nature Publishing Group 2014
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author Mercer, T
Clark, M
Crawford, J
Brunck, M
Gerhardt, D
Taft, R
Nielsen, L
Nielsen, L
Dinger, M
Mattick, J
Mattick, J
author_facet Mercer, T
Clark, M
Crawford, J
Brunck, M
Gerhardt, D
Taft, R
Nielsen, L
Nielsen, L
Dinger, M
Mattick, J
Mattick, J
author_sort Mercer, T
collection OXFORD
description RNA sequencing (RNAseq) samples the majority of expressed genes infrequently, owing to the large size, complex splicing and wide dynamic range of eukaryotic transcriptomes. This results in sparse sequencing coverage that can hinder robust isoform assembly and quantification. RNA capture sequencing (CaptureSeq) addresses this challenge by using oligonucleotide probes to capture selected genes or regions of interest for targeted sequencing. Targeted RNAseq provides enhanced coverage for sensitive gene discovery, robust transcript assembly and accurate gene quantification. Here we describe a detailed protocol for all stages of RNA CaptureSeq, from initial probe design considerations and capture of targeted genes to final assembly and quantification of captured transcripts. Initial probe design and final analysis can take less than 1 d, whereas the central experimental capture stage requires ∼7 d.
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spelling oxford-uuid:808d8a1d-f111-41a4-87b6-7fe8d3c8a1a92022-03-26T21:24:09ZTargeted sequencing for gene discovery and quantification using RNA CaptureSeq.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:808d8a1d-f111-41a4-87b6-7fe8d3c8a1a9EnglishSymplectic Elements at OxfordNature Publishing Group2014Mercer, TClark, MCrawford, JBrunck, MGerhardt, DTaft, RNielsen, LNielsen, LDinger, MMattick, JMattick, JRNA sequencing (RNAseq) samples the majority of expressed genes infrequently, owing to the large size, complex splicing and wide dynamic range of eukaryotic transcriptomes. This results in sparse sequencing coverage that can hinder robust isoform assembly and quantification. RNA capture sequencing (CaptureSeq) addresses this challenge by using oligonucleotide probes to capture selected genes or regions of interest for targeted sequencing. Targeted RNAseq provides enhanced coverage for sensitive gene discovery, robust transcript assembly and accurate gene quantification. Here we describe a detailed protocol for all stages of RNA CaptureSeq, from initial probe design considerations and capture of targeted genes to final assembly and quantification of captured transcripts. Initial probe design and final analysis can take less than 1 d, whereas the central experimental capture stage requires ∼7 d.
spellingShingle Mercer, T
Clark, M
Crawford, J
Brunck, M
Gerhardt, D
Taft, R
Nielsen, L
Nielsen, L
Dinger, M
Mattick, J
Mattick, J
Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title_full Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title_fullStr Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title_full_unstemmed Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title_short Targeted sequencing for gene discovery and quantification using RNA CaptureSeq.
title_sort targeted sequencing for gene discovery and quantification using rna captureseq
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