Samtools mpileup vs bcftools mpileup

Samtools mpileup vs bcftools mpileup. bam| bcftools call -mv -Oz -o calls. One of the things that got dropped in mpileup was the ability to add a consensus call. Please use bcftools mpileup for Feb 4, 2021 · At a position, read maximally 'INT' reads per input file. bcftools mpileup -Q 30 -q 30 -f . Feb 16, 2021 · The first step, initially “samtools mpileup” but subsequently moved to “bcftools mpileup,” reads the alignments and for each position of the genome constructs a vertical slice across all reads covering the position (“pileup”). bai index using samtools. bam -f ref. I already tried to change the default parameters and the best result I can get is less than 50% of the depth observed with samtools-0. What could be the reason Hi all, I read through samtools manuals several times, but I'm still not clear on how exactly samtools & bcftools decide to call a SNP. Please use bcftools mpileup for Jan 14, 2019 · yes, in the mpileup step, the pre-filtering removes reads from PL calculation. fa alignments. Generate text pileup output for one or multiple BAM files. bam, and 3. You are using the parameters . However, it has now been overtaken by GATK HaplotypeCaller 2 (Fig. [mpileup] 1 samples in 1 input files. i. 9) has been removed. Feb 16, 2021 · SAMtools and BCFtools are widely used programs for processing and analysing high-throughput sequencing data. That depends what column it is you need. Killed: 9. Sep 25, 2020 · When I mpileup it again with the following commands bcftools mpileup -C50 -B -Q 0 , I still couldn't see any records in the output. (That branch will work with the samtools/bcftools 1. Do the first pass on variant calling by counting read coverage with bcftools. Each input file produces a separate group of pileup columns in the output. I am using mpileup to generate the counts of each allele for an individual at given loci, command below. And since mpileup is run the same way in both runs, I can think of two possible causes only: either there is some slowdown on the computing cluster unrelated to bcftools, or there is a specific site where the program gets stuck. Aug 29, 2023 · The somatic. You signed out in another tab or window. Comparative Jan 9, 2024 · I checked if there are reads in the region and there are (below is a screenshot of the output of samtools view) Also, bam files and reference use the same chromosome convention name. fna -b bam_list. It does a map-reduce using temporary files. Jul 5, 2022 · In particular, Samtools mpileup (now Bcftools mpileup) was previously the most widely used variant caller 1. It seems horribly inefficient to run the same command twice instead of reusing the file obtained from previous step (especially because generating mpileup Oct 25, 2015 · This command will parallelize over chromosomes/contigs with one simultaneous job per core, writing all results to my. # SAMTools mpileup # -b List of BAM files # -f Reference FASTA file # -l Use positions in BED file # -u Generate uncompressed BCF file # # BCFTools view # -b Output BCF # -e Likelihood based analyses # -c SNP calling # -g Call genotypes at variant sites # -v Output potential variant sites only Jul 5, 2022 · Identification of genetic variations is a central part of population and quantitative genomics studies based on high-throughput sequencing data. 6. Jan 27, 2020 · Bcftools mpileup should be used instead of samtools mpileup for variant calling. bcf # filter adjacent indels within 5bp bcftools filter --IndelGap 5 calls. In versions of samtools <= 0. fa - > output ", while the command for mpileup is "samtools mpileup -uf hg19. " DESCRIPTION. in *samtools mpileup* the default was highly likely to be increased and the. bam | bcftools view -bvcg - > var. raw. pjotrp self-assigned this on Mar 10, 2015. 1. bcftools mpileup -Ou -f reference. So in fact it is @gringer's suggestion that is instructions for an earlier version of samtools / bcftools. bam, I can either run samtools mpileup 1. sorted. It thinks whatever you are putting after -R is a file and tries to open it. 17) indicates that the output option -U, mwu-u will revert the new tags (with Z) to the previous format (without Z). Field values are always displayed before tag values. samtools mpileup -f reference. bam as the output. 19 calling was done with bcftools view. Jan 31, 2019 · Then I wanted to use mpileup for variant calling and used this command:. bcftools Jul 5, 2022 · These results suggest that Bcftools mpileup may be the first choice for non-human studies and that variants within repeats might have to be excluded for downstream analyses. It imports from and exports to the SAM (Sequence Alignment/Map) format, does sorting, merging and indexing, and allows to retrieve reads in any regions swiftly. $\endgroup$ – Mar 9, 2015 · Sambamba mpileup is somewhat experimental and not well tested. Multiple samples: Aug 22, 2019 · The mapping qualities given in sam files are different to those in the mpileup output. fasta \. gz bcftools index calls. Most BCFtools commands accept the -i, --include and -e, --exclude options which allow advanced filtering. This will be most effective on a cluster, so as to spread the IO load. 3), I do not seem to be able to set ploidy to 1. Here we will give some examples on how you can do so with bcftools. pileup) was only 2 kb while that of SRA Oct 31, 2018 · You need a file name after the -R option (for example, a BED file with the regions you want). answered Jul 26, 2022 at 13:59. bam, 3. bam > trio. I just followed 'Manual Reference Pages - samtools', my command line is like this; samtools mpileup -C50 -gf ref. 3. It can help understanding what complex tools and pipelines actually do. fa bbm. bam That's what I see: [warning] samtools mpileup option v is functional, but deprecated. the reference genome must be passed. fasta myData. Aug 2, 2018 · The samtools mpileup and bcftools mpileup should give about the same result. It still uses an astonishing amount of memory to parse your headers, which may or may not be practical to improve. Filtering. vcf Oct 16, 2020 · 2023/07/24 mpileup修正. flt-indels. It includes programs for performing variant calling (mpileup-bcftools). I would like to ask regarding a value reported in vcf output after SNP calling process through mpileup. dad. bam and then samtools mpileup 2. * bcftools (when used) If --samtools is skipped, samtools mpileup is called with default arguments ´samtools´. jkbonfield/htslib. It looks like a gap from 106943 to 108043. The two indels share the same reads and after realignment end up as essentially the same call, just one base apart. fa Jun 15, 2021 · SAMtools is a suite of commands for dealing with databases of mapped reads. Sambamba mpileup relies on external tools and acts as a multi-core implementation of samtools + bcftools. Users are now required to choose between the old samtools calling model (-c/--consensus-caller) and the new multiallelic calling model (-m/--multiallelic-caller). I'm not sure why - it's useful, and one of the reasons for later adding samtools consensus. pileup. All commands work transparently with both VCFs and BCFs, both uncompressed and BGZF-compressed. fa calls. We then pipe the output to bcftools, which does our SNP calling based on those likelihoods. When I replace this command by bcftools mpileup I lose most of the reads. . The mapping score is a Phred-scaled probability of unique mapping, by definition. I think it seems like the variant frequency of base type in a certain . In the second calling step these are used to jointly call variants Apr 18, 2016 · See bcftools call for variant calling from the output of the samtools mpileup command. Bcftools mpileup replaces the samtools one, and I think bcftools consensus should be able to generate a fastq from the VCF. bam file and its . Bcftools mpileup uses mapping scores to evaluate variant calling in a way that the variant calling score of an SNV is not allowed to be higher than the mapping score. Here is my command line and the associated error: "| bcftools call --ploidy 1 -mv >", vcf_file)) Jan 13, 2021 · I have seen piping samtools with varscan as follows. The mpileup still ran though but the file size (SRA. It's unclear to me when this difference in tags was introduced. The consensus is written either as FASTA, FASTQ, or a pileup oriented format. bam []] Input options: -R, --regions-file FILE restrict to regions listed in a file. This will result in 7 reads got to mpileup. So you skip all reads with mapping quality <20. ) Aug 22, 2021 · This suggests that there is no significant difference between running bcftools call with and without -C,-T. vcf. By the way, the version of bcftools is v1. Samtools mpileup VCF and BCF output (deprecated in release 1. I was able to do the alignment using BWA and thus obtain a . fa. bcftools is itself a comprehensive pipeline and produces a variant call format (VCF) that is used in many downstream analyses. 8 participants. When executing bcftools call on the output of bcftools mpileup it sometimes fails to retain deletions with approriate coverage. bam, 2. samtools mpileup --output-extra FLAG,QNAME,RG,NM in. Using bcftools/1. fa --bam-list slice --annotate AD Feb 28, 2019 · I am using samtools mpileup for snp calling. Where my_bams. So I'm still having trouble with the output bcf file Sep 29, 2020 · DP is the raw read depth, AD includes only high-quality bases. My next step is to perform variant calling using bcftools. vcf -v file-sorted. If your organism has 20 chromosomes, submit 20 jobs to your cluster, each doing 'samtools mpileup' on a different chromosome. bam mom. int rflag_skip_any_unset, rflag_skip_all_unset, rflag_skip_any_set, rflag_skip_all_set, output_type; You signed in with another tab or window. @pd3 is the person to confirm. Oct 21, 2021 · The missingness (of variants) for samples is around ~60%. jar mpileup2snp. fofn -r {1} :::: genome. bam SamTools: Mpileup¶ SamToolsMpileup · 1 contributor · 2 versions. vcf" the two sample code is "bcftools mpileup -Ou -b bam. (#1584 Jul 14, 2023 · 1. In the first step (the mpileup step ), we process the reads, identify likely alleles, and compute genotype likelihoods. Thanks again. vcf After filtering, kept 3 out of 3 Individuals After filtering, kept 14117 out of a possible 14117 Sites #now quality filter the raw calls bcftools filter --exclude 'QUAL < 30' raw_calls. Suppose we have reference sequences in ref. I am writing a code to analyse yeast sequencing data using R. •Popular tools include Samtools and GATK (from Broad) •Germline vs Somatic mutations •Samtools: Samtools’s mpileup (formerly pileup) computes genotype likelihoods supported by the aligned reads (BAM file) and stores in binary call format (BCF) file. Mpileup: Input: BAM file Output: Pileuped up reads under the reference. 17 c350a57 - well before any of the current maintainers started working on samtools. Reload to refresh your session. BCFtools uses two algorithms. 33. Feb 16, 2021 · bcftools mpileup \ -r chrM \ --output-type v \ --fasta-ref "${fasta_filename}" \ --max-depth 8000 \ --skip-indels \ ${bam_filenames} [mpileup] maximum number of reads per input file set to -d 8000 [mplp_func] Skipping because 2756366 is outside of 16571 [ref:24] [mplp_func] Skipping because 2781409 is outside of 16571 [ref:24] [mplp_func] Skipping because 2804105 is outside of 16571 [ref:24 First let's see how to use a simple pipeline to identify genetic variants using bcftools mpileup and bcftools call. fa, indexed by samtools faidx, and position sorted alignment files aln1. Note that the original samtools mpileup command had a minimum value of 8000/n where n was the number of input files given to mpileup. txt > raw. fa aln1. Aug 26, 2015 · There are various tools for variant detection on Sanger sequences but I wanted to take this opportunity to check out SAMtools mpileup and BCFtools. Samtools mpileup can still produce VCF and BCF output (with -g or -u), but this feature is deprecated and will be removed in a future release. Generate consensus from a SAM, BAM or CRAM file based on the contents of the alignment records. We are using a number of non-default options: Estimating genotype likelihoods with bcftools mpileup: -q 20: filter out sites with base quality (BQ) <20 samtools mpileup --output-extra FLAG,QNAME,RG,NM in. Users are now required to choose between the old samtools calling model (-c/--consensus-caller) and the new multiallelic calling model (-m/--multiallelic-caller). fai > my. As an alternative, try bcftools mpileup -x -B -m3 -h500 to disable both BAQ and overlap removal (plus some saner default values). It starts at the first base on the first chromosome for which there is coverage and prints out one line per base. fas HA1. gz -Ob -o calls. Best if you post the two lines including any output from the commands. It's a simple parseable Aug 4, 2020 · bcftools mpileup -Ou -f ref. bam aln2. The original samtools package has been split into three separate but tightly coordinated projects: htslib: C-library for handling high-throughput sequencing data; samtools: mpileup and other tools for handling SAM, BAM, CRAM; bcftools: calling and other tools for handling VCF, BCF pileup意为“堆叠”，就如同其名字一样， samtools mpileup 命令可以在给定的基因组区间内的每个碱基位置（column）上将mapping到这个位置上的所有reads堆叠集中起来，得到输入BAM在这个给定的基因组区间内每个碱基位置也即每个column上的整体信息。. e. bcftools mpileup -Ob -o output. will display four extra columns in the mpileup output, the first being a list of comma-separated read names, followed by a list of flag values, a list of RG tag values and a list of NM tag values. Author. Feb 6, 2012 · i reinstalled ubuntu and i installed samtools by downloading from sourceforge. These match the equivalent long options found in "samtools mpileup" and gives a consistent way of specifying the base and mapping quality filters. $ bcftools mpileup. We will use the command mpileup. bcf -Ob -o calls. /samtools mpileup -f file. -f ref. Samtools is designed to work on a stream. 9, we have been having an issue when trying to pileup the first position in a contig. I've tried to run through multiple combination of arguments with mpileup (-B, -C, -q, etc) & bcftools, but still ran into the problem below. Apr 5, 2023 · The old pileup was removed years ago in 0. fasta sample1. bam, the following command lines call SNPs and short INDELs: samtools mpileup -uf ref. --output-sep CHAR. Whenever I use samtools mpileup -uf pfal. bam sample2. Variant calling. bam | java -jar VarScan. Indexing the reference… again. If strict thresholds are applied and an indel is not considered for calling, and also no SNP is present, a site will appear as identical to reference. The limit can be changed by running eg ulimit -n 9999. The multiallelic calling Nov 21, 2017 · #use vcftools to get information about our variant set vcftools --vcf raw_calls. pjotrp added the bug label on Mar 10, 2015. 1 ), partly because Samtools calculates the genotype likelihoods. norm. But when I look the bam file in IGV, I can see the mapped reads and the variants in the 'missing' region. gz bcftools mpileup on its own runs well on the same samples. samtools mpileup -f Spombe_genome. the math is a bit more complicated than just picking the highest For this tutorial, we will use bcftools which is designed by the same team behind samtools - they are part of the same pipeline. bcf -f ref. Bcftools is for example used in Snippy the variant calling and core genome alignment sowftware that is implemented in ALPPACA Jul 7, 2022 · Image from “Data Wrangling and Processing for Genomics”. bam | bcftools call -c > bbm. samtools mpileup -B -q 1 \. You switched accounts on another tab or window. 多様なコマンドから成る。. My command in pileup is "samtools view -u Inputfile (BAM) | samtools pileup -vcf hg19. Feb 18, 2013 · First, samtools mpileup command transposes the mapped data in a sorted BAM file fully to genome-centric coordinates. fna. My question is that what is the meaning of a value called "QS", which states "Auxiliary tag used for calling" from its description. pileup: parallel --colsep '\t' samtools mpileup -b my_bams. Furthermore you are skipping bases with base quality <20. Please use bcftools mpileup instead. pl being a perl script probably means it'll work fine on the modern bcftools VCF, but I've never used it and I don't know how it differs to bcftools consensus itself. If you are running a reasonably recent version of bcftools, this is described in the header. gz or any mpileup command I am getting [E::faidx_fetch_seq] The sequence "MN918127" not bcftools call 中有 -c -m 参数. The default output for FASTA and FASTQ formats include one base per non-gap consensus. May 13, 2019 · Try to run ulimit -n to see if there is a system limit on the number of simultaneously open files. #Run VarScan trio. bcftoolsは変異をコールして バリアントコールフォーマットの VCFを出力したり、VCFやBCF（VCFのバイナリーフォーマット）を操作するツール。. Hello there, I am using samtools mpileup for snp calling. /ref/scaffold. , if I have 3 bam files, 1. py script expects INFO tags RPB, MQB, BQB, and MQSB ( lines 106-109 ). the original *samtools mpileup* command had a minimum value of '8000/n'. bam | bcftools call -vmO z --format-fields GQ,GP -o output. The cram files were generated from exome array, and the average genotyping rate should be above 98%. Here are two examples. Mar 12, 2012 · No matter I used pileup or mpileup, the problem is always there. Make mpileup's overlap removal choose a random sequence. Even though popular variant callers such as Bcftools mpileup and GATK HaplotypeCaller were developed nearly 10 years ago, their performance is still largely unknown for non-human species. This means that. vcf or any mpileup command I am getting [E::faidx_adjust_position] The sequence "Pf3D7_01_v3 | organism=Plasmodium_falciparum_3D7 | version=2015-06-18 | length=640851 | SO=chromosome" not found for all position. bam Variant Calling using Samtools (Mpileup + bcftools)¶ Samtools calculates the genotype likelihoods. I won't have time to look into it before April. In order to avoid tedious repetion, throughout this document we will use "VCF" and "BCF" interchangeably, unless I am curious if it matters whether or not it matters if I run mpileup individually for each bam file versus running mpileup as one process for all the bam files. bam child. Jan 11, 2022 · Alignment and sorting the file by Coordinate using STAR, Duplication removal with Picard, samtools sort for the output file resulted from the Picard's step and samtools index accordingly, and. bcftools: Input: Pileup output from Mpileup Output: VCF file with sites and genotypes See bcftools call for variant calling from the output of the samtools mpileup command. (The "Source code" downloads are generated by GitHub and are incomplete as they don't bundle HTSlib and are missing some generated files. bam [in2. 8 pileup uses the follwing command. Whenever I use bcftools mpileup -Ou -f HA1. fai is the output of samtools faidx or alternately a newline Dec 18, 2018 · That's right, thanks very much for the bug report. In this post, I illustrate the BWA-MEM, SAMtools mpileup, and BCFtools pipeline with a bunch of randomly generated sequences. flie -f ref. Learning Objectives Sep 14, 2021 · The problem is, BCFTools expects mpileup to be piped from another command and I cannot find an input parameter to specify a mpileup file that already exists (the one used with VarScan2). I guess samtools is doing some recalculation. This algorithm reads the alignments and, for each position of the genome, constructs a vertical slice across all reads samtools mpileup -f [reference sequence] [parameters] [BAM file(s)] >myData. I don't # call variants bcftools mpileup -Ou -f reference. tar. vcf" the different result show you in blow: sample1 result: #CHROM POS ID REF ALT QUAL FILTER INFO FORMAT fatherHap # call variants bcftools mpileup -Ou -f reference. samtoolsの論文で発表された（論文より "The SAMtools package consists of two key Apr 2, 2022 · Today I'm trying to clarify my understanding of --max-depth argument from bcftools mpileup command, which is currently described as: At a position, read maximally INT reads per input file. lomereiter added the in progress label on Apr 10, 2015. This is the official development repository for samtools. Interestingly, bcftools mpileup documentation (version 1. The first thing we need to do is index our reference Apr 22, 2020 · DarioS commented on Apr 22, 2020. ) Changes affecting the whole of bcftools, or multiple commands: The -i / -e filtering expressions. 1 设置基因组 Development. To avoid generating intermediate temporary files, the output of bcftools mpileup is piped to bcftools call. bam. New work and changes: Added --min-BQ and --min-MQ options to depth. makes the actual call. 3. Therefore, the following tools should be present in the PATH: * samtools. You can debug the problem by leaving out the bcftools call command for now and check the difference in the outputs. Dec 31, 2015 · When using Version: 1. fasta -s sample1,sample2 sample1. Since cram files don't contain the reference file, I was just wondering if the bcftools call didn't call the variant if the site stays the same with See bcftools call for variant calling from the output of the samtools mpileup command. The -m switch tells the program to use the default calling method, the -v option asks to output only variant sites, finally the -O option E. In the examples below, we demonstrate the usage on the query command because it allows us to show the output in a very compact form using the -f formatting option. Sep 19, 2014 · Samtools is a set of utilities that manipulate alignments in the BAM format. Note that. The overall genotyping rate is ~0. Please switch to using bcftools mpileup in future. 1 tags/releases. May 14, 2012 · The simplest way to do this is to divide the work up by reference sequence. Please use bcftools mpileup for Second, bcftools call identifies both variants and genotypes, i. 8 pileup. 15 (C) Adam Auton and Anthony Marcketta 2009 Parameters as interpreted: --vcf raw_calls. 17: Download the source code here: bcftools-1. fa BAM | bcftools view -bvcg -> output". where 'n' was the number of input files given to mpileup. The variant calling command in its simplest form is. fai -b bam_list. fasta aln. bcftools: Input: Pileup output from Mpileup Output: VCF file with sites and genotypes. With neither of them you're likely to get the right answer assuming your depth is high (which it almost certainly is for Covid-19 samtools mpileup --output-extra FLAG,QNAME,RG,NM in. This is selected using the -f FORMAT option. fa | bcftools call -mv -o sample1. Hands-on: Step 1: Calculate the read coverage of positions in the genome. fa -r chr2:1 test. bcf. You'll be using it quite a bit throughout the course. fa Jul 8, 2019 · $\begingroup$ The VCF/BCF-outputting mpileup command was transferred from samtools to bcftools relatively recently. -R is defined as "As for -r, --regions, but regions read from FILE; see Common Options" The definition of -r shows you can use the format chr1:12345 or chr2, for example. Findings. fa | bcftools call -mv -o sample1and2. Because mpileup does not keep track of reads used for indel calling between positions, both indel variants are reported. As this suggests the process has two steps. Mar 23, 2021 · the one sample code is "bcftools mpileup -Ou sample1. 3 (using htslib 1. Usage: bcftools mpileup [options] in1. #Generate a three-sample mpileup. However, when I click on the Common Options hyperlink, I see "FILE Files can be both VCF or BCF, uncompressed or BGZF-compressed. 理解是 -c 用来call 普通二倍体 单样品;-m 可以call 多样品合并的bam ,发现一些稀有变异,应该就是 群call. If that's not possible because of permissions (bash: ulimit: open files: cannot modify limit: Operation not permitted), edit the following files and add these lines: Apr 26, 2016 · Try compiling both samtools and bcftools against htslib's hugeheaders branch (PR samtools/htslib#373); this should suffice to fix this issue. The first one is called mpileup. mpileup. Samtools mpileup however has two different formats with the default always being a simple columnar format showing chr, pos, reference, depth, base-calls and qualities. Sep 25, 2018 · The update worked fine for all the steps expect the one using samtools-0. fofn is a file of BAM files, and genome. This works as expected: $ bcftools mpileup -f test. (Of course the old vcfutils. My command is below. This is fixed now. Note, to save disk space and file I/O, you can redirect mpileup output directly to VarScan with a “pipe” command. E. bam > file1 (it's compressed binary Mar 11, 2024 · BCFtools is a widely-used variant calling tool, especially among non-human species, which is characterized by its small time of execution and its precision. The second call part makes the actual calls. bcftools utilities are assotiated or part of the Samtools suite of programs. 2015-01-12. vcf #returns this: VCFtools - 0. 双等位基因 (biallelic sites )和多等位基因 (multiallelic sites) 序言：七十年代末，一起剥皮案震惊了整个滨河市，随后出现的几起案子，更 May 15, 2020 · Saved searches Use saved searches to filter your results more quickly Jan 4, 2022 · OmBibo commented on Jan 4, 2022. net (latest version) as you know, 'pileup' option is deprecated and replaced with 'mpileup' option. This tutorial expects you have already completed the Mapping tutorial. Regards, Dan. g. Nov 2, 2018 · Looking in the code, this seems to be an ugly bug caused by mpileup's site-oriented variant calling. If --bcftools is used without parameters, samtools is May 21, 2013 · SAMtools is a suite of commands for dealing with databases of mapped reads. SamTools: Mpileup¶ SamToolsMpileup · 1 contributor · 2 versions. bcftools release 1. But how to pipe the following pipeline for varscan trio. Bcftools applies the priors (from above) and calls variants (SNPs and indels). Apr 22, 2021 · See also samtools/htslib#1273, which is vital for calling on amplicon sequencing. 17. bz2 . bam, etc or run samptools mpileup 1. primer trimming with ivar, with SRA_clean. Jul 5, 2022 · Bcftools mpileup uses alignments of a mapper as it is. That is, the VCF / BCF output mode of mpileup is better in bcftools. I Dec 17, 2010 · The basic Command line. Successfully merging a pull request may close this issue. gz # normalize indels bcftools norm -f reference. bam >/dev/null; echo $? [mpileup] 1 samples in 1 input files 0 Bu BCFtools is a program for variant calling and manipulating files in the Variant Call Format (VCF) and its binary counterpart BCF. They include tools for file format conversion and manipulation, sorting, querying, statistics, variant calling, and effect analysis amongst other methods. fa bams/M3*realigned. bam | bcftools call -mv -Oz -o calls. The first mpileup part generates genotype likelihoods at each genomic position with coverage. bcf # apply variants to create consensus sequence cat reference. *-d* parameter would have an effect only once above Generate text pileup output for one or multiple BAM files. bam and aln2. bam (I know Jul 25, 2022 · The problem was that although an index must have been build, in the following code instead of passing the index, bcftools mpileup -Ou -f index. One sample: samtools mpileup -f reference. The INFO/DP tag displays the number of raw reads. fasta -o test. fv xt ul mo lo eg ku zz jp cz