Hi! I want to use Beagle version 5.1. to exclusively impute my missing genotypes (no ref. panel needed). I used .ped and .map files to create my .vcf file using plink1.09b. My file 394.RMV-UNKNOW-SEXUAL-Filtered.vcf looks like:
contig=<ID=29,length=51502869>
INFO=<ID=PR,Number=0,Type=Flag,Description="Provisional reference allele, may not be based on real refere$
FORMAT=<ID=GT,Number=1,Type=String,Description="Genotype">
CHROM POS ID REF ALT QUAL FILTER INFO FORMAT 1_V_UK561500501863_plate1_A01 2_V$
1 16947 BovineHD0100000005 C T . . PR GT 0/1 0/0 0/0
1 135098 Hapmap43437-BTA-101 T C . . PR GT 0/1 0/1 0/0
1 149772 BovineHD0100000042 G A . . PR GT 0/0 0/0 0/1
1 158820 BovineHD0100000048 G T . . PR GT 0/1 1/1 0/0
1 163995 BovineHD0100000051 T C . . PR GT 0/0 0/0 0/0
1 183040 BovineHD0100000057 T G . . PR GT 0/1 0/1 ./.
1 267940 ARS-BFGL-NGS-16466 T C . . PR GT 0/1 0/0 1/1
1 290690 BovineHD0100000082 A C . . PR GT 0/0 0/1 0/1
When I run the command:
java -Xss5m -Xmx4g -jar /exports/eddie/scratch/v1mmart8/Test/beagle.18May20.d20.jar gt=394.RMV-UNKNOW-SEXUAL-Filtered.vcf out=394_imputed impute=false
The output created 394_imputed.vcf.gz is empty. The error file says_ERROR: REF field is not a sequence of A, C, T, G, or N characters at 1:78986953 [D]
I tried to include missing=./. in the command but beagle does not recognize it.
Any idea why is it not running? Many thanks!
Hi! Many thanks for your response!
As you said, many issues here: 1)With egrep '78986953' I get 1 78986953 ARS-BFGL-NGS-119431 D I . . PR GT 0/0 I think the D is the problem here.. I made a quick search and this is the only case where this happens so I think I can solve it by removing this line. 2)Also, about the missings, I have them coded like "." for single characters in the columns of quality and filter, which is "./." when occurs in a SNP. However, when I include the command missing=. in Beagle I get the error: Error: unrecognized parameter: missing=. I wonder how Beagle wants the missing values to be coded? 3) I also detected I have some repeated SNPs (by position) in my .vcf file, any idea how to remove them?
To jean.elbers: no, I just want to impute my missing SNPs in my 50k file of 386 animals
Many thanks!
I have removed the line with "D", and now beagle runs but does not finishes the work. I have aprox. 36 000 SNPs but it stops at line 568 and the error: WARNING: We recommend that you use a minimum of 4 GB of virtual memory when running Java 1.8.0_74 on Eddie. Please see the following for details: https://www.wiki.ed.ac.uk/display/ResearchServices/Java Exception in thread "main" java.lang.IllegalArgumentException: Duplicate marker: 1 59409838 ARS-USMARC-Parent-DQ404150-rs29012530_dup T C at vcf.Markers.markerSet(Markers.java:131) at vcf.Markers.<init>(Markers.java:85) at vcf.Markers.create(Markers.java:64) at vcf.BasicGT.markers(BasicGT.java:105) at vcf.BasicGT.<init>(BasicGT.java:86) at vcf.TargetData.targGT(TargetData.java:92) at vcf.TargetData.advanceWindowCm(TargetData.java:120) at main.Main.phaseData(Main.java:158) at main.Main.main(Main.java:113)
Some help to interpret it would be really useful Many thanks! Marina
change -Xmx4g to -Xmx100g or however much ram you have available (use free -h)
Hi, many thanks for the reply! I tried several but I am restricted to 4g.. (-Xmx4g). However, I think my issue is that I have duplicated SNPs in my .vcf file.. Any ideas how to delete them? Many thanks!
See how to remove duplicate SNP rows in vcf using bcftools norm, I guess that is what you want?
Dear Jean,
Thanks, I follow it and it worked!. Just a last question so I make sure I did not miss anything Do we use the same commands (call java, gt= and out=) to phase a reference database with Beagle and to impute missing values in our case study population without a reference? (of course with different entry and ouput files) Is it normal that I do not get any imputation accuracy if I am only imputing missing positions in the genotyped SNPs? Many thanks for your time!
Please see http://faculty.washington.edu/browning/beagle/beagle_5.1_08Nov19.pdf especially section 5 about output
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I might be wrong but I did not see the option
missing. Also should your REF field contain missing genotypes? I think you have a malformed VCF file, or theIDfield contains some special character that is causing that error. Can you find1:78986953in the vcf file and post it? The other issue I noticed is that your missing data is.when I think it should be./..Are you trying to impute 3 samples without a reference panel? I do not think this will work well at all. I would think more like 100-1000 might be okay. The more the better without a reference panel.