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Transcript Assembly Merge
#CUFFMERGE Use Cuffmerge to merge predicted transcripts from all libraries into a unified transcriptome Refer to the Cufflinks manual for a more detailed explanation: http://cufflinks.cbcb.umd.edu/manual.html#cuffmerge
Cuffmerge basic usage cuffmerge [options]* <assembly_GTF_list.txt> <assembly_GTF_list.txt> is a text file "manifest" with a list (one per line) of GTF files that you would like to merge together into a single GTF file. Extra options specified below '-p 8' tells cuffmerge to use eight CPUs '-o' tells cuffmerge to write output to a particular directory '-g' tells cuffmerge where to find reference gene annotations. It will use these annotations to gracefully merge novel isoforms (for de novo runs) and known isoforms and maximize overall assembly quality. '-s' tells cuffmerge where to find the reference genome files
Merge all 4 cufflinks results so that they will have the same set of transcripts for comparison purposes
For reference guided mode: cd $RNA_HOME/expression/tophat_cufflinks/ref_guided/ ls -1 cDNAlib*/transcripts.gtf > assembly_GTF_list.txt cat assembly_GTF_list.txt cuffmerge -p 8 -o merged -g $RNA_HOME/refs/hg19/genes/genes_chr22.gtf -s $RNA_HOME/refs/hg19/bwt/22/ assembly_GTF_list.txt
For de novo mode: cd $RNA_HOME/expression/tophat_cufflinks/de_novo/ ls -1 cDNAlib*/transcripts.gtf > assembly_GTF_list.txt cat assembly_GTF_list.txt cuffmerge -p 8 -o merged -g $RNA_HOME/refs/hg19/genes/genes_chr22.gtf -s $RNA_HOME/refs/hg19/bwt/22/ assembly_GTF_list.txt
NOTICE: This resource has been moved to rnabio.org. The version here will be maintained for legacy use only. All future development and maintenance will occur only at rnabio.org. Please proceed to rnabio.org for the current version of this course.
Table of Contents
Module 0: Authors | Citation | Syntax | Intro to AWS | Log into AWS | Unix | Environment | Resources
Module 1: Installation | Reference Genomes | Annotations | Indexing | Data | Data QC
Module 2: Adapter Trim | Alignment | IGV | Alignment Visualization | Alignment QC
Module 3: Expression | Differential Expression | DE Visualization
Module 4: Alignment Free - Kallisto
Module 5: Ref Guided | De novo | Merging | Differential Splicing | Splicing Visualization
Module 6: Trinity
Module 7: Trinotate
Appendix: Saving Results | Abbreviations | Lectures | Practical Exercise Solutions | Integrated Assignment | Proposed Improvements | AWS Setup