# HG changeset patch # User gregory-minevich # Date 1349717910 14400 # Node ID 41803b10e893e06ee42daa0bc794e112964faa15 # Parent bdaadaee1cbb581ba0c83e7c923e510b24829194 Uploaded diff -r bdaadaee1cbb -r 41803b10e893 SNP_Mapping.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/SNP_Mapping.xml Mon Oct 08 13:38:30 2012 -0400 @@ -0,0 +1,143 @@ + + Map a mutation by plotting recombination frequencies resulting from crossing to a highly polymorphic strain + + #if $source.source_select=="elegans" #SNP_Mapping.py --sample_vcf $sample_vcf --loess_span $loess_span --d_yaxis $d_yaxis --h_yaxis $h_yaxis --points_color "$points_color" --loess_color "$loess_color" --output $output --location_plot_output $location_plot_output --standardize $standardize --normalize_bins $normalize_bins --break_file $source.Celegans + #else if $source.source_select=="arabadopsis" #SNP_Mapping.py --sample_vcf $sample_vcf --loess_span $loess_span --d_yaxis $d_yaxis --h_yaxis $h_yaxis --points_color "$points_color" --loess_color "$loess_color" --output $output --location_plot_output $location_plot_output --standardize $standardize --normalize_bins $normalize_bins --break_file $source.Arabadop + #else if $source.source_select=="other" #SNP_Mapping.py --sample_vcf $sample_vcf --loess_span $loess_span --d_yaxis $d_yaxis --h_yaxis $h_yaxis --points_color "$points_color" --loess_color "$loess_color" --output $output --location_plot_output $location_plot_output --standardize $standardize --normalize_bins $normalize_bins --break_file $source.Other + #end if + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + sys + optparse + csv + re + decimal + rpy + + + + + + + +**What it does:** + +This tool is part of the CloudMap pipeline for analysis of mutant genome sequences. For further details, please see `Gregory Minevich, Danny S. Park, Daniel Blankenberg, Richard J. Poole and Oliver Hobert. CloudMap: A Cloud-based Pipeline for Analysis of Mutant Genome Sequences. (Genetics 2012 In Press)`__ + + .. __: http://biochemistry.hs.columbia.edu/labs/hobert/literature.html + +CloudMap workflows, shared histories and reference datasets are available at the `CloudMap Galaxy page`__ + + .. __: http://usegalaxy.org/cloudmap + +This tool improves upon, and automates, the method described in Doitsidou et al., PLoS One 2010 for mapping causal mutations using whole genome sequencing data. + +Sample CloudMap output for a linked chromosome: + +.. image:: http://biochemistry.hs.columbia.edu/labs/hobert/CloudMap/Linked_LG_500px.png + + +The polymorphic Hawaiian strain CB4856 is used as a mapping strain in most cases but in principle any sequenced nematode strain that is significantly different from the mutant strain can be used for mapping. The tool plots the ratio of mapping strain (Hawaiian)/mutant strain (N2) nucleotides at all SNP positions, reflecting the number of recombinants in the sequenced pool of animals. Chromosomes which contain regions of linkage to the causal mutation will have regions where the ratio of mapping strain (Hawaiian)/total reads will be equal to 0. The scatter plots for such linked regions will have a high number of data points lying exactly on the X axis. A loess regression line is plotted through all the points on a given chromosome giving further accuracy to the linked region. + +Each scatter plot has a corresponding frequency plot that displays regions of linked chromosomes where pure parental (mutant strain) alleles are concentrated. 1Mb bins for the 0 ratio SNP positions are colored gray by default and .5Mb bins are colored in red. By default, frequency plots of pure parental alleles are normalized to remove false linkage caused by previously described (Seidel et al. 2008) patterns of genetic incompatibility between Bristol and Hawaiian strains. This normalization can be turned off via a checkbox input form setting. + + +The experimental design required to generate data for the plots is described in the CloudMap paper (Fig.6A): + +.. image:: http://biochemistry.hs.columbia.edu/labs/hobert/CloudMap/Doitsidou_2010_PLoS_Fig.1_500px.png + + +------ + +**Input:** + + +This tool accepts as input a single VCF file containing reference (e.g. Bristol) and alternate (e.g. Hawaiian) mapping strain alleles calls at each of the mapping strain variant positions (e.g. 112,000 Hawaiian SNPs) in the pooled mutant sample. This input VCF is generated at an earlier analysis step by running the GATK Unified Genotyper on a BAM alignment file of the pooled mutant sample with a provided reference file of mapping strain variants (e.g. Hawaiian SNPs) in VCF format. The reader is referred to the user guide and online video for direction on this procedure. + +Default GATK Unified Genotyper parameters for mapping quality, base quality and coverage at each SNP position typically yield good results, though users may experiment with adjusting these parameters. In our testing, low threshold filtering on base pair quality (default settings) has been useful in improving accuracy of plots while high threshold filtering on coverage has skewed plot accuracy. + +The required VCF of mapping strain (e.g. Hawaiian) SNPs is a reference file that contains mapping strain SNP positions and reference base pairs at each position. It is available in the `CloudMap Shared Data library`__ + + .. __: http://usegalaxy.org/library + +You may also make your own VCF of mapping strain variant positions following the steps described in the CloudMap paper. + +The CloudMap Hawaiian Variant Mapping with WGS Data tool supports data from any organism that has been crossed to a mapping strain for which variant information is available. C. elegans and Arabidopsis are natively supported. For all other organisms, users must provide a simple tab-delimited configuration file containing chromosome numbers and respective lengths (example configuration files for most major organisms provided at http://usegalaxy.org/cloudmap). Additional files required for other organisms are the same as described for C. elegans: a VCF file consisting of pooled F2 mutant progeny sequencing data, and a VCF file of the mapping strain variants. + + +**Output:** + +The tool also provides a tabular output file that contains a count of the number of reference and alternate variants at each mapping strain variant position as well as the ratio of mapping strain (e.g. Hawaiian)/alternate SNPs. The position of each mapping strain SNP in map units and physical coordinates is also provided in the output file. + + +------ + +**Settings:** + +.. class:: infomark + +Information on loess regression and the loess span parameter: +http://en.wikipedia.org/wiki/Local_regression + +.. class:: infomark + +Based on our testing, we've settled on .1 as a loess span default. Larger values result in smoothing of the line to reflect trends at a more macro level. Smaller values result in loess lines that more closely reflect local data fluctuations. + +.. class:: infomark + +Supported colors for data points and loess regression line: + +http://www.stat.columbia.edu/~tzheng/files/Rcolor.pdf + +http://research.stowers-institute.org/efg/R/Color/Chart/ColorChart.pdf + + + +.. class:: warningmark + +This tool requires that the statistical programming environment R has been installed on the system hosting Galaxy (http://www.r-project.org/). If you are running this tool on Galaxy via the Cloud, this does not apply to you. + + +------ + +**Citation:** + +This tool is part of the CloudMap package from the Hobert Lab. If you use this tool, please cite `Gregory Minevich, Danny S Park, Daniel Blankenberg, Richard J. Poole, and Oliver Hobert. CloudMap: A Cloud-based Pipeline for Analysis of Mutant Genome Sequences. (Genetics 2012 In Press)`__ + + .. __: http://biochemistry.hs.columbia.edu/labs/hobert/literature.html + +Correspondence to gm2123@columbia.edu (G.M.) or or38@columbia.edu (O.H.) + + +