This package simulates whole genome sequencing using 10X Genomics Linked Read technology. We have attempted to realistically capture all of the relevant steps of the 10X protocol so that it can be used to faithfully evaluate linked read sequencing of different genomes, mutation rates, input libraries, and short read sequencing conditions in silico. We have tested the package with LongRanger, SuperNova, HapCUT2, fragScaff and ARCS to confirm that variant identifation, phasing, de novo assembly and scaffolding are supported. We also encourage users to use these simulations to aid in the development of novel algorithms. Please feel free to contact us if your pipelines require additional features.
git clone --recursive https://github.com/aquaskyline/LRSIM.git
cd LRSIM
sh make.sh
cd test
sh test.sh
- Please review the 10X Genomics website for an overivew of the sequencing process and the definitions of the related terms. Note that 'Molecule' and 'Partition' are synonymous to 'Fragment' and 'Pool'.
- If you encounter the "Missing Inline::C library" error, please install the Inline::C perl library using CPAN or uncomment the following two lines of code by removing the hash symbol at the front.
#use lib "./lib";
#use lib dirname($0)."/lib";
- The simulated reads were tested to be compaitible with "longRanger wgs" and "supernova".
- The default parameters are similar to 10X Chromium's standard protocal for human genome.
- Set -z to run DWGSIM in parallel. For the human genome, each copy of DWGSIM takes 4GB memory. Set -z to the number of available cores if you have enough memory.
- For the human genome, the memory consumption peaks at 48GB, and takes about 5 hours to finish with default parameters.
- With the same output prefix
-p, you can continue from step 4: Simulate reads using option-u 4with different-f(fragment size),-t(partitions to generate) and-m(average number of molecules per partition). This shortern the simulation from 5 hours to 1.5 hours for human. - Please use this pipeline for non-human genomes at your own risk. You may want to use
-oto skip valid range check on parameters. You shouldn't set-mto over 4700, which is the number of available barcodes, or the program will not run to the end. Note that the default barcoding parameters do not perform well for small genomes (<100Mbp). - I hate asking users to install dependencies, so they are included in the repo (make sure to use git clone --recursive). If you still run into problem, please write to me.
- To simulate reads using known variants, please provide LRSIM with variant inserted haploid FASTA files using
-g, separated by comma. I suggest using vcf2diploid to generate haploid FASTAs from VCF. - User can provide ia real fragment size distribution using
-c. A sample file is attest/fragmentSizesList.
Usage: ./simulateLinkedReads.pl -r/-g <reference/haplotypes> -p <output prefix> [options]
Reference genome and variants:
-d INT Haplotypes to simulate [2]
-g STRING Haploid FASTAs separated by comma. Overrides -r and -d.
-1 INT 1 SNP per INT base pairs [1000]
-2 INT Minimum length of Indels [1]
-3 INT Maximum length of Indels [50]
-4 INT # of Indels [1000]
-5 INT Minimum length of Duplications and Inversions [1000]
-6 INT Maximum length of Duplications and Inversions [10000]
-7 INT # of Duplications and # of Inversions [100]
-8 INT Minimum length of Translocations [1000]
-9 INT Maximum length of Translocations [10000]
-0 INT # of Translocations [100]
Illumina reads characteristics:
-e FLOAT Per base error rate of the first read [0.0001,0.0016]
-E FLOAT Per base error rate of the second read [0.0001,0.0016]
-i INT Outer distance between the two ends for pairs [350]
-s INT Standard deviation of the distance for pairs [35]
Linked reads parameters:
-b STRING Barcodes list
-x INT # million reads pairs in total to simulated [600]
-f INT Mean molecule length in kbp [100]
-c STRING Input a list of fragment sizes. Overrrides -f.
-t INT n*1000 partitions to generate [1500]
-m INT Average # of molecules per partition [10]
Miscellaneous:
-u INT Continue from a step [auto]
1. Variant simulation
2. Build fasta index
3. DWGSIM
4. Simulate reads
5. Sort reads extraction manifest
6. Extract reads
-z INT # of threads to run DWGSIM [8]
-o Disable parameter checking
-h Show this help
The ratio of homozygous to heterzygous simulated variant is hardcoded as 1:2.
The simulator uses a modified version of DWGSIM originally developed by Nils Homer (nh13/DWGSIM) and SURVIVOR by Fritz Sadlezeck (fritzsedlazeck/SURVIVOR).
The MIT License (MIT)
Copyright (c) 2016 Ruibang Luo <[email protected]>
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