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De Novo Plant Transcriptome Assembly and Annotation Using Illumina RNA-Seq Reads

  • Stephanie C. Kerr
  • Federico Gaiti
  • Milos TanurdzicEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1933)

Abstract

The ability to identify and quantify transcribed sequences from a multitude of organisms using high-throughput RNA sequencing has revolutionized our understanding of genetics and plant biology. However, a number of computational tools used in these analyses still require a reference genome sequence, something that is seldom available for non-model organisms. Computational tools employing de Bruijn graphs to reconstruct full-length transcripts from short sequence reads allow for de novo transcriptome assembly. Here we provide detailed methods for generating and annotating de novo transcriptome assembly from plant RNA-seq data.

Key words

RNA-seq Long noncoding RNA Trinity De novo transcriptome assembly 

Supplementary material

418724_1_En_16_MOESM1_ESM.pl (1 kb)
Supplementary File 1 : The get_longest_ORF_per_transcript.pl Perl script used in Subheading 3.5, step 5 for identifying the longest open reading frame from each transcript present in your transcriptome assembly (PL 1 kb)
418724_1_En_16_MOESM2_ESM.sh (4 kb)
Supplementary File 2 : The lncRNA_pipeline.sh bash script used in Subheading 3.5, step 9 (SH 4 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Stephanie C. Kerr
    • 1
  • Federico Gaiti
    • 2
  • Milos Tanurdzic
    • 1
    Email author
  1. 1.School of Biological SciencesThe University of QueenslandSt LuciaAustralia
  2. 2.New York Genome Center and Department of MedicineWeill Cornell MedicineNew YorkUSA

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