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 The input for the FLUX CAPACITOR is the annotation of a reference transcriptome and reads fromRNAseq technologies aligned to the genome. From the reference annotation, splicing graphs are produced and reads are mapped to corresponding edges in these graphs according to the position where they align in the genomic sequence. The resulting graph with edges labelled by the number of reads can be interpreted as a flow network where each transcript representing a transportation path from its start to its end and consequently each edge a possibly shared segment of transportation along which a certain number of reads per nucleotide -- i.e., a flux -- is observed. Given a density function of reads along a transcript, the expected participation of each transcript in an edge under consideration can be estimated. The basic idea is to cast back from these latter participations and the observed number of reads - allowing for a certain amount of noise - to the original transcript abundancies. To do so, a linear constraint is formalized for each edge, and an optimal solution for the complete set of constraints is found by a standard linear program solver. 

 

The basic problem addressed by the FLUX CAPACITOR.

The exonic structure of two spliceforms (labeled as "SF A" and "SF B") is shown, with aligned reads from by RNAseq methods (top) . Those reads mapped to the edges of a splicing graph (bottom) represent a signal, measured as the FLUX - the relative coverage along an exonic stretch. Where transcripts overlap in exons, their respective flux is combined. Given the information from all edges in a locus, signal separation is achieved by decomposition across a flow

 

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Page: 1 - Introduction
Background The input for the FLUX CAPACITOR is the annotation of a reference transcriptome and reads fromRNAseq technologies aligned to the genome. From the reference annotation, splicing graphs are produced and reads are mapped to corresponding edges in
Page: 1.0.1 Release Notes
Page: 1.0.2 Release Notes
Page: 1.1 Release Notes
Page: 1.2 Release Notes
Page: 1.2.1 Release Notes
Page: 1.2.2 Release Notes
Page: 1.2.3 Release Notes
Page: 1.2.4 Release Notes
Page: 1.3 Release Notes
Page: 1.4 Release Notes
Page: 1.5.1 Release Notes
Release 1.5.1 is a direct replacement of release 1.5, containing a hotfix for the bug below.
Page: 1.5.2 Release Notes
Page: 1.6 Release Notes
Page: 1.6.1 Release Notes
Page: 2 - Download
For bug reporting and questions please see Appendix C - JIRA Issue Tracker. Stable Versions Date Download Release notes 2014-04-09 Flux Capacitor 1.6.1 http://artifactory.sammeth.net/artifactory/barna/barna/barna.capacitor/1.6.1/flux-capacitor-1.6.1.tgz 1
Page: 3 - Getting Started
Page: 3.1 - System Requirements
Java Virtual Machine The Flux Capacitor is written in 100% pure Java, therefore no platform-specific compiling of the bytecode is required. A Java Virtual Machine (JVM) with level 1.6 or higher has to be installed on the system, however, it is recommended
Page: 3.2 - Starting the Flux Capacitor
Start Command The Flux Capacitor provides a wrapper script to launch the program from predominant shell interpreters of major platforms. shell (Linux/Unix) $ flux-capacitor cmd (Windows) C:\>flux-capacitor.bat -t capacitor When started with the --help fl
Page: 3.3 - Examples
GTEx runs Flux Version Flux-Capacitor v1.2.3 (Flux Library: 1.20) Annotation File The quantifications are based on a modified Gencode (v12) version created at the Broad Institute: http://www.broadinstitute.org/cancer/cga/tools/rnaseqc/examples/gencode.v12
Page: 4 - Annotation Mapping
Subsections
Page: 4.1 - Segment Graph
Similar to the concept of splicing graphs [Heber 2002 http://bioinformatics.oxfordjournals.org/content/18/suppl_1/S181.abstract], we employ a graph structure for representing the reference transcriptome that is quantified in a non-redundant data structure
Page: 4.2 - Read Mapping
To avoid redundancy caused by overlapping exons of alternative transcripts, we employ read mappings to the genome. However, our data structure also permits mappings to de novo transcriptome assemblies given that one provides coordinates relative to the pr
Page: 5 - Deconvolution
Subsections
Page: 5.1 - Linear Program
We interpret G with each edge colored by read mappings as a flow network, considering the read volume assigned to every (super-) edge as a flux created by the expression of the underlying supporting transcripts T.Consequently, given an edge the contributi
Page: 5.2 - Bias estimation
RNA-Seq is known to carry inherit biases from the experimental setup, convoluted effects of enzymatic preferences in particular steps of the applied protocol, for instance fragmentation, reverse transcription and adapter ligation. Models of deconvolution
Page: 6 - Quantified Elements
Subsections
Page: 6.3 Intron Quantifications
List of features that are quantified by the Flux, and the corresponding attributes that are provided in the ouput. Transcripts transcript_id gene_id / locus_id nr. reads length RPKM Splice Junctions gene_id/locus_id nr.reads The coordinates (start,end)

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Page: Analysis of Geuvadis Dataset using Flux-Capacitor.
Hi all, Hopefully this is the correct place to ask this question. I'm looking to use flux-capacitor to reference the Geuvadis dataset against gencodeV18.gtf. I want to keep the methodology exactly the same as was done in the original mapping, changing onl
Page: Appendix A - File Format Specifications
Page: Appendix B - Frequently Asked Questions (FAQ)
Page: Appendix C - JIRA Issue Tracker
The software company Atlassian http://www.atlassian.com/ granted to the Flux Capacitor project an Open Source License for their JIRA tracking system http://sammeth.net/jira. Please feel free to employ this system to report any issue (e.g., bugs, requests
Page: Appendix D - Forum
The Flux Capacitor Forum is subdivided into discussion groups:

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Page: BAM - Mapping Locations
SAM (Sequence Alignment/Map) format is a generic format for storing large nucleotide sequence alignments. BAM format is the compressed binary version of SAM format. Further information about these formats can be found in the SAM format specifications http
Page: BED - Insert File
The insert file provides information on the fragments (so-called "inserts") described by the mappings of paired-end read data. Inserts are reported after annotation mapping, with their insert sizes according to the procecessed (spliced) transcript coordin

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Page: Cant find tool capacitor
I'm having problems starting Flux-capacitor. I have the correct java RunTim env: C:\>java -version java version "1.6.0_20" Java (TM) SE Runtime Environment (build 1.6.0_20-b02) Java HotSpot(TM) Client VM (build 16.3-b01, mixed mode) When I launch the wra

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Page: D.1 - FAQ (Frequently Asked Questions)
Doubts that arise regularly when using the Flux Capacitor. File here any questions of general/technical nature, but remember that reports about errors in the program should be reported as issues in the tracking system instead, to be handled by a developer
Page: D.2 - Annotation Mapping
Discussions about mapping reads that have been aligned to a reference genome to a corresponding transcript annotation. This section provides room for threads about the rules that assign a read or read-pair to a certain exon, exon/exon- or splice-junction.
Page: D.3 - Bias Estimation
Discussions about approaches to estimate read distributions from experimental datasets.
Page: D.4 - Deconvolution
Discussions about predicting transcript abundances.

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Page: Error while iterating loci and [ERROR] java.lang.ArrayIndexOutOfBoundsException: 0
Hi, I just run Flux Capacitor using parameter.txt: ANNOTATION_FILE HoSa_build37.2_Genes_genes.gtf COUNT_ELEMENTS [SPLICE_JUNCTIONS, INTRONS] ANNOTATION_MAPPING AUTO ./flux-capacitor -p parameter.txt -i control.sorted.bam -o control.fluxcapacitor.gtf &

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Page: Flux Capacitor output files
I am sorry, but I do not seem to be able to find any documentation describing the output files. The standart output (gtf format) is easy, but What is in each column of the coverage file and the insert file? Thank you, Ana.
Page: Flux Capcacitor error: java.lang.OutOfMemoryError: Java heap space with FLUX_MEM=16G
I have been using Flux Capacitor to quantify transcript abundance for samples in my RNA-seq study. However, for 2 of 126 samples, Flux Capacitor crashes with an out of memory error. I have increased the java heap space to 16G using the environmental varia

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Page: Geuvadis Quantifications
Flux Version Flux Capacitor 1.0.1 http://sammeth.net/artifactory/gradle-barna/barna/barna.capacitor/1.0.1/flux-capacitor-1.0.1.tgz Read Mappings Unfiltered GEM read mappings were employed, after converting them to BED format. Annotation File The quantific
Page: GTEx Quantifications
Flux Version Flux Capacitor 1.2.3 http://sammeth.net/artifactory/barna/barna/barna.capacitor/1.2.3/flux-capacitor-1.2.3.tgz (Flux Library: 1.20) Read Mappings Tophat mappings were employed, filtered for primary alignments (mapper comparison) by samtools s

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Home page: Home
For reporting bugs please go to the JIRA Issue Tracker, to ask a question visit the Forum.
Page: How to get name and sequence of reads mapped to a particular gene
Hello, I am trying to look at some of the actual reads that mapped to a particular gene using the GEUVADIS data set. For example, lets say I have a bed file for one individual (SAMPLE1.bed) and the gencode annotation file (GENCODE.v12.gtf). I would li
Page: How to quantify multiple bam files
Dear Sammeth, Is there any support to quantify multiple bam files using flux capacitor. I can quantify one file using following command: flux-capacitor -p param.txt -i myMapping1.bam -a myAnnotation.gtf -o myout.flux --threads 2 I mean, is there a
Page: How to quantify the expression value of transcript in RPKM
Dear All, I am trying to measure the expression values ( in RPKM ) for gencode lncRNA uploaded in your site, using flux capacitor. But it shows RPKM 0.000 for all transcript.Can you please suggest a sample command in details? I am using following command

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Page: JSON - Profile File
The profile file gives the following information in json format: 5' to 3' read distribution along transcripts mapping statistics read coverage statistics All the information stored in the profile are derived from single transcript loci only. The read dis

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Page: Overlapping transcripts - RPKM NAN
Dear Sammeth, I run flux-capacitor in the following options: flux-capacitor -i bamfile -a annotation_file -o ouptut --threads nthreads. When I run it with an gff annotation file that have some transcripts that overlap each other, but not exons, such as I

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Page: PAR - Parameters
The PAR format in the Flux Capacitor is used to administrate all parameters of a run. It is a simple format containing key value pairs (one per line) with the following parameter names (i.e., keys): File Locations Key Type Default Value Description ANNOTA
Page: Problems working with BAM files
When running the capacitor with BAM input files, I get the following error: [ERROR] Exception creating BAM index for record IPMHD:09324:09018 78b aligned read. net.sf.samtools.SAMException: Exception creating BAM index for record IPMHD:09324:09018 78b ali

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Page: Read Descriptors
The BARNA Descriptor The BARNA (Barcelona Attributes for RNA-Seq) descriptor is a proposition for annotating unique read IDs with additional information yielded from special experiments, e.g., read mates derived from the same cDNA molecule, information ab

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Page: Understanding RPKM values
Hi, I ran flux capacitor on a human BAM file with 9902168 aligned paired end reads (19804336 aligned reads which equals the number of SAM records as I consider primary alignments only); I used the parameter file ANNOTATION_FILE ensembl_human_71.gtf CO

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Page: You cannot specify a READ_Descriptor for BAM files
Hi, I'm annotating some filtered, sorted bam files with their index files and when I run: flux-capacitor -p <parameter_file> -i <sample.bam> -o <output.gtf> with: ANNOTATION_FILE gencode.v12.annotation.gtf READ_DESCRIPTOR {ID}/{MATE}[1,2] ANNOTATION_MAP

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