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− | (''WARNING: Under Construction'')
| + | '''Note:''' the latest version of this practical is available at: [[SeqShop: Genetic Association Analysis Practical]] |
| + | * The ones here is the original one from the June workshop (updated to be run from elsewhere) |
| + | |
| + | |
| + | == Introduction == |
| + | [[Media:Seqshop association 2014 06.pdf|View Lecture Slides]] |
| + | |
| + | [[Media:Seqshop association practice 2014 06.pdf|View Introductory Slides for Practical Session]] |
| | | |
| == Goals of This Session == | | == Goals of This Session == |
| + | |
| * Understand how to annotate variants using EPACTS | | * Understand how to annotate variants using EPACTS |
| * Understand how to run single variant association analysis using EPACTS | | * Understand how to run single variant association analysis using EPACTS |
| * Understand how to run rare variant association test using EPACTS | | * Understand how to run rare variant association test using EPACTS |
| * Understand how to visualize the association output from EPACTS | | * Understand how to visualize the association output from EPACTS |
| + | |
| + | == Setup in person at the SeqShop Workshop == |
| + | ''This section is specifically for the SeqShop Workshop computers.'' |
| + | <div class="mw-collapsible mw-collapsed" style="width:600px"> |
| + | ''If you are not running during the SeqShop Workshop, please skip this section.'' |
| + | <div class="mw-collapsible-content"> |
| | | |
| {{SeqShopLogin}} | | {{SeqShopLogin}} |
| | | |
− | == Setup your run environment== | + | === Setup your run environment=== |
− | | + | This is the same setup you did for the previous tutorial, but you need to redo it each time you log in. |
− | This is a setup similar to the Structural Variant session. | |
| | | |
− | * GotCloud program | + | This will setup some environment variables to point you to |
| + | * [[GotCloud]] program |
| * Tutorial input files | | * Tutorial input files |
| * Setup an output directory | | * Setup an output directory |
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| * You won't see any output after running <code>source</code> | | * You won't see any output after running <code>source</code> |
| ** It silently sets up your environment | | ** It silently sets up your environment |
− | ** If you want to view the detail of the set up, type | + | ** If you want to view the detail of the setup, type |
− | less /home/hmkang/seqshop/setup.txt | + | less /home/mktrost/seqshop/setup.txt |
| and press 'q' to finish. | | and press 'q' to finish. |
| + | |
| <div class="mw-collapsible mw-collapsed" style="width:200px"> | | <div class="mw-collapsible mw-collapsed" style="width:200px"> |
| View setup.txt | | View setup.txt |
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| export SV=/home/hmkang/seqshop/reference/svtoolkit | | export SV=/home/hmkang/seqshop/reference/svtoolkit |
| export EXT=/home/hmkang/seqshop/external | | export EXT=/home/hmkang/seqshop/external |
| + | export EPACTS=/home/hmkang/seqshop/epacts |
| export OUT=~/out | | export OUT=~/out |
| mkdir -p ${OUT} | | mkdir -p ${OUT} |
| + | </div> |
| + | </div> |
| + | </div> |
| + | </div> |
| + | |
| + | |
| + | == Setup when running on your own outside of the SeqShop Workshop == |
| + | ''This section is specifically for running on your own outside of the SeqShop Workshop.'' |
| + | <div class="mw-collapsible" style="width:600px"> |
| + | ''If you are running during the SeqShop Workshop, please skip this section.'' |
| + | <div class="mw-collapsible-content"> |
| + | |
| + | This tutorial builds on the alignment & snpcall tutorials, if you have not already, please first run those tutorials: [[SeqShop:_Sequence_Mapping_and_Assembly_Practical, June 2014|Alignment Tutorial]] & [[SeqShop: Variant Calling and Filtering for SNPs Practical, June 2014|SNP Calling Tutorial]] |
| + | |
| + | === Download & Build EPACTS === |
| + | If you do not already have EPACTS: |
| + | * cd to where you want EPACTS installed (you can change this to any directory you want) |
| + | mkdir -p ~/seqshop |
| + | cd ~/seqshop/ |
| + | * download, decompress, and build the version of epacts that was tested with this tutorial: |
| + | wget http://csg.sph.umich.edu//kang/epacts/download/EPACTS-3.2.6.tar.gz |
| + | tar xvf EPACTS-3.2.6.tar.gz |
| + | cd EPACTS-3.2.6 |
| + | ./configure --prefix=$HOME/seqshop/epacts |
| + | make |
| + | make install |
| + | cd ../.. |
| + | |
| + | {{SeqShopRemoteEnv}} |
| + | |
| + | <ul> |
| + | <li> Additional variables for EPACTS:</li> |
| + | <ul> |
| + | <div class="mw-collapsible" style="width:500px"> |
| + | <li>Using bash (replace the paths below with the appropriate paths):</li> |
| + | <div class="mw-collapsible-content"> |
| + | :<pre>export EPACTS=~/seqshop/epacts</pre> |
| + | </div> |
| + | </div> |
| + | <div class="mw-collapsible mw-collapsed" style="width:500px"> |
| + | <li>Using tcsh (replace the paths below with the appropriate paths):</li> |
| + | <div class="mw-collapsible-content"> |
| + | :<pre>setenv EPACTS ~/seqshop/epacts</pre> |
| + | </div> |
| + | </div> |
| + | </ul> |
| + | </ul> |
| + | |
| </div> | | </div> |
| </div> | | </div> |
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| Check the contents of the VCF file using the following command. | | Check the contents of the VCF file using the following command. |
| | | |
− | zless $OUT/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz | + | zless ${OUT}/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz |
| | | |
| === Phenotype Information === | | === Phenotype Information === |
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| Phenotype information is prepared in PED format commonly used in other GWAS software such as MERLIN or PLINK. | | Phenotype information is prepared in PED format commonly used in other GWAS software such as MERLIN or PLINK. |
| | | |
− | less $IN/assoc/seqshop.ped | + | less ${SS}/assoc/seqshop.ped |
| | | |
| The first several line should look like below. | | The first several line should look like below. |
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| View Data | | View Data |
| <div class="mw-collapsible-content" style="width:800px"> | | <div class="mw-collapsible-content" style="width:800px"> |
− | #FAM_ID IND_ID DAD_ID MOM_ID SEX GENO PHENO | + | #FAM_ID IND_ID DAD_ID MOM_ID SEX PHENO |
− | HG00551 HG00551 0 0 0 1 0 | + | HG00551 HG00551 0 0 0 0 |
− | HG00553 HG00553 0 0 0 0 0 | + | HG00553 HG00553 0 0 0 0 |
− | HG00554 HG00554 0 0 0 0 0 | + | HG00554 HG00554 0 0 0 0 |
− | HG00637 HG00637 0 0 0 0 0 | + | HG00637 HG00637 0 0 0 0 |
− | HG00638 HG00638 0 0 0 0 0 | + | HG00638 HG00638 0 0 0 0 |
− | HG00640 HG00640 0 0 0 1 1 | + | HG00640 HG00640 0 0 0 1 |
− | HG00641 HG00641 0 0 0 2 1 | + | HG00641 HG00641 0 0 0 1 |
− | HG00734 HG00734 0 0 0 1 1 | + | HG00734 HG00734 0 0 0 1 |
− | HG00736 HG00736 0 0 0 0 0 | + | HG00736 HG00736 0 0 0 0 |
− | HG00737 HG00737 0 0 0 0 0
| |
| ... | | ... |
| </div> | | </div> |
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| | | |
| mkdir --p $OUT/assoc | | mkdir --p $OUT/assoc |
− | $EPACTS/bin/epacts-anno --in $OUT/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz --out $OUT/assoc/snps.anno.vcf.gz | + | $EPACTS/bin/epacts-anno --in $OUT/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz --out $OUT/assoc/snps.anno.vcf.gz --ref $SS/ref22/human.g1k.v37.chr22.fa |
| | | |
| Then you will see a series of messages before annotation finishes. | | Then you will see a series of messages before annotation finishes. |
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| <div class="mw-collapsible-content" style="width:800px"> | | <div class="mw-collapsible-content" style="width:800px"> |
| /home/hmkang/seqshop/epacts/bin/anno -i /net/seqshop-server/hmkang/out/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz -r \ | | /home/hmkang/seqshop/epacts/bin/anno -i /net/seqshop-server/hmkang/out/thunder/chr22/ALL/thunder/chr22.filtered.PASS.beagled.ALL.thunder.vcf.gz -r \ |
− | /home/hmkang/seqshop/epacts/share/EPACTS/human_g1k_v37.fasta -f refGene -g /home/hmkang/seqshop/epacts/share/EPACTS/hg19_gencodeV14.txt.gz \ | + | /home/hmkang/seqshop/ref22/human_g1k_v37.chr22.fa -f refGene -g /home/hmkang/seqshop/epacts/share/EPACTS/hg19_gencodeV14.txt.gz \ |
| -c /home/hmkang/seqshop/epacts/share/EPACTS/codon.txt -o /net/seqshop-server/hmkang/out/assoc/snps.anno.vcf.gz --inputFormat vcf \ | | -c /home/hmkang/seqshop/epacts/share/EPACTS/codon.txt -o /net/seqshop-server/hmkang/out/assoc/snps.anno.vcf.gz --inputFormat vcf \ |
| -p /home/hmkang/seqshop/epacts/share/EPACTS/priority.txt | | -p /home/hmkang/seqshop/epacts/share/EPACTS/priority.txt |
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| Gene Annotation | | Gene Annotation |
| Parameters : -g [/home/hmkang/seqshop/epacts/share/EPACTS/hg19_gencodeV14.txt.gz] | | Parameters : -g [/home/hmkang/seqshop/epacts/share/EPACTS/hg19_gencodeV14.txt.gz] |
− | -r [/home/hmkang/seqshop/epacts/share/EPACTS/human_g1k_v37.fasta] | + | -r [/home/hmkang/seqshop/ref22/human_g1k_v37.chr22.fa] |
| --inputFormat [vcf], --checkReference, -f [refGene] | | --inputFormat [vcf], --checkReference, -f [refGene] |
| -p [/home/hmkang/seqshop/epacts/share/EPACTS/priority.txt] | | -p [/home/hmkang/seqshop/epacts/share/EPACTS/priority.txt] |
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| -u [], -d [], --se [], --si [], --outputFormat [] | | -u [], -d [], --se [], --si [], --outputFormat [] |
| Other Annotation Tools : --genomeScore [], --bed [], --tabix [] | | Other Annotation Tools : --genomeScore [], --bed [], --tabix [] |
− | Load reference genome /home/hmkang/seqshop/epacts/share/EPACTS/human_g1k_v37.fasta... | + | Load reference genome /home/hmkang/seqshop/ref22/human_g1k_v37.chr22.fa... |
− | DONE: 84 chromosomes and 3101804739 bases are loaded. | + | DONE: 1 chromosomes and 51304566 bases are loaded. |
| Load codon file /home/hmkang/seqshop/epacts/share/EPACTS/codon.txt... | | Load codon file /home/hmkang/seqshop/epacts/share/EPACTS/codon.txt... |
| DONE: codon file loaded. | | DONE: codon file loaded. |
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| * How can you represent the variant in terms of amino acid changes? | | * How can you represent the variant in terms of amino acid changes? |
| | | |
− | === Run Single Variant Analysis === | + | == Single Variant Association Analysis == |
| + | |
| + | Let's run a single-variant association analysis using a score test. |
| + | |
| + | $EPACTS/bin/epacts-single --ped $SS/assoc/seqshop.ped --vcf $OUT/assoc/snps.anno.vcf.gz --out $OUT/assoc/single --region 22:36000000-37000000 --test b.score --pheno PHENO --run 2 |
| + | |
| + | After running it, you will see EPACTS output files by looking at |
| + | |
| + | ls $OUT/assoc |
| + | |
| + | The top association results can be viewed by |
| + | |
| + | head $OUT/assoc/single.epacts.top5000 |
| + | |
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View top association results |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | #CHROM BEGIN END MARKER_ID NS AC CALLRATE MAF PVALUE SCORE NS.CASE NS.CTRL AF.CASE AF.CTRL |
| + | 22 36995620 36995620 22:36995620_A/G 62 36 1 0.29032 5.6717e-09 5.8262 31 31 0.51613 0.064516 |
| + | 22 36993088 36993088 22:36993088_G/C 62 30 1 0.24194 7.3258e-07 4.9525 31 31 0.43548 0.048387 |
| + | 22 36997871 36997871 22:36997871_G/T 62 30 1 0.24194 7.3258e-07 4.9525 31 31 0.43548 0.048387 |
| + | 22 36987368 36987368 22:36987368_G/A 62 31 1 0.25 2.0898e-06 4.7445 31 31 0.43548 0.064516 |
| + | 22 36987861 36987861 22:36987861_A/G 62 31 1 0.25 2.0898e-06 4.7445 31 31 0.43548 0.064516 |
| + | 22 36985499 36985499 22:36985499_C/T 62 29 1 0.23387 5.7389e-06 4.5358 31 31 0.40323 0.064516 |
| + | 22 36978260 36978260 22:36978260_G/T 62 28 1 0.22581 1.5051e-05 4.3279 31 31 0.3871 0.064516 |
| + | 22 36998907 36998907 22:36998907_C/T 62 61 1 0.49194 0.00015557 -3.782 31 31 0.30645 0.67742 |
| + | 22 36667082 36667082 22:36667082_T/G 62 28 1 0.22581 0.0003506 -3.5747 31 31 0.080645 0.37097 |
| + | </div> |
| + | </div> |
| + | |
| + | You can look also visualize the results by QQ-plot and Manhattan plot |
| + | |
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View QQ plots |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | :<pre>evince $OUT/assoc/single.epacts.qq.pdf&</pre> |
| + | [[File:Single.epacts.qq.png]] |
| + | </div> |
| + | </div> |
| | | |
− | * Run score test
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View Manhattan plots |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | :<pre>evince $OUT/assoc/single.epacts.mh.pdf&</pre> |
| + | [[File:Single.epacts.mh.png|900px]] |
| + | </div> |
| + | </div> |
| | | |
− | mkdir ~/out/assoc
| + | Also, you can create a zoom plot focusing on the region of interest |
− | $S5/epacts/bin/epacts single --ped $S5/examples/index/chr7.CFTR.ped --vcf ~/out/snps/beagle/chr7/chr7.filtered.PASS.beagled.vcf.gz --pheno PHENO --out ~/out/assoc/single.b.score --test b.score --anno --ref $S5/examples/chr7Ref/hs37d5.chr7.fa --region 7:117000000-117500000 --run 1
| |
| | | |
− | * Look at the association statistics
| + | $EPACTS/bin/epacts-zoom --vcf $OUT/assoc/snps.anno.vcf.gz --pos 22:36995620 --prefix $OUT/assoc/single |
| | | |
− | head ~/out/assoc/single.b.score.epacts.top5000
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View Zoom Plots |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | :<pre>evince $OUT/assoc/single.zoom.22.36995620.pdf&</pre> |
| + | [[File:Single.zoom.22.36995620.png]] |
| + | </div> |
| + | </div> |
| | | |
− | * Variant annotation with EPACTS
| + | If you want to run EMMAX, you first need to create a kinship matrix |
| | | |
− | $S5/epacts/bin/epacts anno --in ~/out/snps/beagle/chr7/chr7.filtered.PASS.beagled.vcf.gz --out ~/out/snps/chr7.filtered.PASS.beagled.anno.vcf.gz --ref $S5/examples/chr7Ref/hs37d5.chr7.fa | + | $EPACTS/bin/epacts-make-kin --vcf $OUT/assoc/snps.anno.vcf.gz --min-maf 0.01 --out $OUT/assoc/snps.anno.kinf --run 2 --chr 22 |
| | | |
− | === Visualizing Zoom Plot ===
| + | And run EMMAX test specifying the kinship matrix |
| | | |
− | * Use ENCODE GM12878 cell type to diplsy variants in the locusZoom-like plot
| + | $EPACTS/bin/epacts-single --ped $SS/assoc/seqshop.ped --vcf $OUT/assoc/snps.anno.vcf.gz --out $OUT/assoc/emmax --region 22:36000000-37000000 --test q.emmax --pheno PHENO --run 2 --kinf $OUT/assoc/snps.anno.kinf |
| | | |
− | $S5/epacts/bin/epacts-zoom --vcf ~/out/snps/chr7.filtered.PASS.beagled.anno.vcf.gz --pos 7:117149147 --prefix ~/out/assoc/single.b.score --cellType Gm12878
| + | Then the results may look similar to previous ones. |
| | | |
− | Copy the output file single.b.score.zoom.7.117149147.pdf using WinSCP or other file transfer software
| + | cat $OUT/assoc/emmax.epacts.top5000 |
| | | |
− | === Run Groupwise Test ===
| + | == Run Groupwise Test == |
| | | |
− | * Create a set of markers to test
| + | To run group-wise test such as gene-level burden test, you need to create a marker group file. If your VCF is already annotated, you can create a group file by running |
| | | |
− | $S5/epacts/bin/epacts make-group --vcf ~/out/snps/chr7.filtered.PASS.beagled.anno.vcf.gz --out ~/out/snps/chr7.filtered.PASS.beagled.anno.grp --nonsyn | + | $EPACTS/bin/epacts make-group --vcf $OUT/assoc/snps.anno.vcf.gz --out $OUT/assoc/snps.anno.grp --nonsyn |
| | | |
− | * Run SKAT-O test
| + | The group file is simply a list of marker per group name, as shown below. |
| | | |
− | $S5/epacts/bin/epacts group --ped $S5/examples/index/chr7.CFTR.ped --vcf ~/out/snps/chr7.filtered.PASS.beagled.anno.vcf.gz --out ~/out/assoc/group.skat.o --groupf ~/out/snps/chr7.filtered.PASS.beagled.anno.grp --test skat --skat-o --run 2 | + | cat $OUT/assoc/snps.anno.grp |
| + | APOL1 22:36655735_G/A 22:36657740_G/A 22:36661330_G/A 22:36661566_G/A 22:36661646_G/A 22:36661891_G/A 22:36661906_A/G |
| + | APOL2 22:36623731_T/C 22:36623920_G/A 22:36629466_T/A 22:36633107_C/A |
| + | APOL3 22:36537763_C/T 22:36537798_G/A 22:36556768_G/A 22:36556823_G/T |
| + | APOL4 22:36587154_G/T 22:36587202_G/A 22:36587223_G/T 22:36587346_C/T 22:36587511_C/T 22:36587704_T/C 22:36587886_C/T 22:36593714_G/A 22:36597744_A/C 22:36598049_C/G 22:36598058_T/C 22:36598081_A/T |
| + | APOL5 22:36122356_G/A 22:36122380_T/A 22:36122930_C/T 22:36123083_C/T 22:36124860_C/G |
| + | FOXRED2 22:36900271_T/C 22:36900806_A/G |
| + | MYH9 22:36681163_G/C 22:36684354_T/C 22:36710183_T/C |
| + | Metazoa_SRP 22:36711990_C/G |
| + | RBFOX2 22:36424450_A/C |
| + | RP4-633O19__A.1 22:36792162_G/A |
| | | |
− | cat ~/out/assoc/group.skat.o.epacts | cut -f 4,6,10,11
| + | If you have your own annotation, you can create your own burden test unit by modifying this file. |
| | | |
− | * Run Variable Threshold test
| + | If you want to run a collapsing burden test (CMC), run the following command |
− | $S5/epacts/bin/epacts group --ped $S5/examples/index/chr7.CFTR.ped --vcf ~/out/snps/chr7.filtered.PASS.beagled.anno.vcf.gz --out ~/out/assoc/group.VT --groupf ~/out/snps/chr7.filtered.PASS.beagled.anno.grp --test VT --run 2
| |
| | | |
− | cat ~/out/assoc/group.VT.epacts | cut -f 4,6,10,13 | + | $EPACTS/bin/epacts group --ped $SS/assoc/seqshop.ped --vcf $OUT/assoc/snps.anno.vcf.gz --out $OUT/assoc/group.collapse --test b.collapse --groupf $OUT/assoc/snps.anno.grp --pheno PHENO --run 2 |
| + | |
| + | You can view the results by examining the output file |
| + | cat $OUT/assoc/group.collapse.epacts |
| + | |
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View Output file |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | #CHROM BEGIN END MARKER_ID NS FRAC_WITH_RARE NUM_ALL_VARS NUM_PASS_VARS NUM_SING_VARS PVALUE STATRHO |
| + | 22 36655735 36661906 22:36655735-36661906_APOL1 62 0.14516 7 4 0 0.42748 1 |
| + | 22 36623731 36633107 22:36623731-36633107_APOL2 62 0.064516 4 1 0 0.038657 NA |
| + | 22 36537763 36556823 22:36537763-36556823_APOL3 62 0.080645 4 2 0 0.40634 0 |
| + | 22 36587154 36598081 22:36587154-36598081_APOL4 62 0.14516 12 4 0 0.67891 0 |
| + | 22 36122356 36124860 22:36122356-36124860_APOL5 62 0.1129 5 2 0 0.15055 0.3 |
| + | 22 36900271 36900806 22:36900271-36900806_FOXRED2 NA NA 2 0 0 NA NA |
| + | 22 36681163 36710183 22:36681163-36710183_MYH9 62 0.032258 3 1 0 1 NA |
| + | 22 36711990 36711990 22:36711990-36711990_Metazoa_SRP NA NA 1 0 0 NA NA |
| + | 22 36424450 36424450 22:36424450-36424450_RBFOX2 62 0.032258 1 1 0 1 NA |
| + | 22 36792162 36792162 22:36792162-36792162_RP4-633O19__A.1 NA NA 1 0 0 NA NA |
| + | </div> |
| + | </div> |
| + | |
| + | You can run SKAT-O test in a similar way, but with a special tag |
| + | |
| + | $EPACTS/bin/epacts group --ped $SS/assoc/seqshop.ped --vcf $OUT/assoc/snps.anno.vcf.gz --out $OUT/assoc/group.skato --test skat --skat-o --groupf $OUT/assoc/snps.anno.grp --pheno PHENO --run 2 |
| + | |
| + | And view output files |
| + | |
| + | cat $OUT/assoc/group.skato.epacts |
| + | |
| + | <div class="mw-collapsible mw-collapsed" style="width:400px"> |
| + | View Output file |
| + | <div class="mw-collapsible-content" style="width:800px"> |
| + | #CHROM BEGIN END MARKER_ID NS FRAC_WITH_RARE NUM_ALL_VARS NUM_PASS_VARS NUM_SING_VARS PVALUE STATRHO |
| + | 22 36655735 36661906 22:36655735-36661906_APOL1 62 0.14516 7 4 0 0.42748 1 |
| + | 22 36623731 36633107 22:36623731-36633107_APOL2 62 0.064516 4 1 0 0.038657 NA |
| + | 22 36537763 36556823 22:36537763-36556823_APOL3 62 0.080645 4 2 0 0.40634 0 |
| + | 22 36587154 36598081 22:36587154-36598081_APOL4 62 0.14516 12 4 0 0.67891 0 |
| + | 22 36122356 36124860 22:36122356-36124860_APOL5 62 0.1129 5 2 0 0.15055 0.3 |
| + | 22 36900271 36900806 22:36900271-36900806_FOXRED2 NA NA 2 0 0 NA NA |
| + | 22 36681163 36710183 22:36681163-36710183_MYH9 62 0.032258 3 1 0 1 NA |
| + | 22 36711990 36711990 22:36711990-36711990_Metazoa_SRP NA NA 1 0 0 NA NA |
| + | 22 36424450 36424450 22:36424450-36424450_RBFOX2 62 0.032258 1 1 0 1 NA |
| + | 22 36792162 36792162 22:36792162-36792162_RP4-633O19__A.1 NA NA 1 0 0 NA NA |
| + | </div> |
| + | </div> |