Bioinformatic Analysis of Small RNA Sequencing Libraries.

Bioinformatic analysis of small RNA sequencing libraries consists of transforming a series of small RNA sequencing experiment fastq files into a table containing small RNA sequences and their abundance. This is achieved by cleaning the reads, aligning the cleaned reads to a reference, and parsing the alignment results. In this protocol we present the most common option, and the rationale, for each of these steps.

[1]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[2]  Wing Hung Wong,et al.  SeqMap: mapping massive amount of oligonucleotides to the genome , 2008, Bioinform..

[3]  Marcel Martin,et al.  Atropos: specific, sensitive, and speedy trimming of sequencing reads , 2017, PeerJ.

[4]  Baohong Zhang,et al.  Conservation and divergence of plant microRNA genes. , 2006, The Plant journal : for cell and molecular biology.

[5]  Mark D. Robinson,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[6]  M. Axtell,et al.  Revisiting Criteria for Plant MicroRNA Annotation in the Era of Big Data[OPEN] , 2018, Plant Cell.

[7]  Richard S. Taylor,et al.  MicroRNA annotation of plant genomes − Do it right or not at all , 2017, BioEssays : news and reviews in molecular, cellular and developmental biology.

[8]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration , 2012, Briefings Bioinform..

[9]  Jonathan M. Yeoh,et al.  Improved Placement of Multi-mapping Small RNAs , 2016, G3: Genes, Genomes, Genetics.

[10]  A. Conesa,et al.  Differential expression in RNA-seq: a matter of depth. , 2011, Genome research.

[11]  Jikai Lei,et al.  miR-PREFeR: an accurate, fast and easy-to-use plant miRNA prediction tool using small RNA-Seq data , 2014, Bioinform..

[12]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer , 2011, Nature Biotechnology.

[13]  Robert Gentleman,et al.  Software for Computing and Annotating Genomic Ranges , 2013, PLoS Comput. Biol..

[14]  Philip C J Donoghue,et al.  Evolutionary history of plant microRNAs. , 2014, Trends in plant science.

[15]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[16]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[17]  Gayathri Mahalingam,et al.  Sample sequencing of vascular plants demonstrates widespread conservation and divergence of microRNAs , 2014, Nature Communications.

[18]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[19]  Hong Ma,et al.  Conservation and divergence of small RNA pathways and microRNAs in land plants , 2017, Genome Biology.

[20]  F. Thibaud-Nissen,et al.  Araport11: a complete reannotation of the Arabidopsis thaliana reference genome , 2016, bioRxiv.

[21]  Robert D. Finn,et al.  Rfam 12.0: updates to the RNA families database , 2014, Nucleic Acids Res..

[22]  Zhiping Weng,et al.  DNApi: A De Novo Adapter Prediction Algorithm for Small RNA Sequencing Data , 2016, PloS one.

[23]  John L. Bowman,et al.  Gene regulation: Ancient microRNA target sequences in plants , 2004, Nature.

[24]  Ana Kozomara,et al.  miRBase: annotating high confidence microRNAs using deep sequencing data , 2013, Nucleic Acids Res..

[25]  Peter F. Stadler,et al.  ViennaRNA Package 2.0 , 2011, Algorithms for Molecular Biology.

[26]  Robert D. Finn,et al.  Rfam 13.0: shifting to a genome-centric resource for non-coding RNA families , 2017, Nucleic Acids Res..

[27]  C. Scutt,et al.  The evolutionary-developmental analysis of plant microRNAs , 2010, Philosophical Transactions of the Royal Society B: Biological Sciences.

[28]  M. Axtell,et al.  Genome-wide analysis of single non-templated nucleotides in plant endogenous siRNAs and miRNAs , 2016, bioRxiv.

[29]  Marcel Martin Cutadapt removes adapter sequences from high-throughput sequencing reads , 2011 .