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Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations
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Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

DOI:
10.3791/54090-v

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11:52 min

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August 04, 2016

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, , ,
1Department of Internal Medicine, Division of Medical Oncology, Comprehensive Cancer Center,The Ohio State University, 2Department of Pharmacology,The Ohio State University

Chapters

  • 00:05Title
  • 01:08Hybridization
  • 03:02Target Capture
  • 06:56Post Capture PCR Amplification
  • 09:45Results: The Capture Method Has Multiple Advantages over Non-targeted RNAseq
  • 11:13Conclusion

Summary

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We describe a targeted RNA sequencing-based method that includes preparation of indexed cDNA libraries, hybridization and capture with custom probes and data analysis to interrogate selected transcripts for gene expression, mutations, and gene fusions. Targeted RNAseq permits cost-effective, rapid evaluation of selected transcripts on a desktop sequencer.

Tags

Keywords: Targeted RNA SequencingGene ExpressionGenomic AlterationsRibosomal RNA DepletionChemical FragmentationCDNA SynthesisBarcoded LibrariesHybridizationTarget CaptureMultiplexed SequencingGene FusionsSingle Nucleotide VariationsAlternative SplicingCustom ProbesCot-1 DNABlocking OligosHybridization BufferStreptavidin Beads

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