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archive:bioinformatic_tools:soapdenovo [2010/04/14 10:44]
galt
archive:bioinformatic_tools:soapdenovo [2015/07/28 06:26] (current)
ceisenhart ↷ Page moved from bioinformatic_tools:soapdenovo to archive:bioinformatic_tools:soapdenovo
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 ==== Overview ==== ==== Overview ====
  
-SOAP = Short_Oligonucleotide_Analysis_Package\\+SOAP = **S**hort **O**ligonucleotide **A**nalysis **P**ackage\\
 SOAPdenovo assembles short oligonucleotide into contigs and scaffolds SOAPdenovo assembles short oligonucleotide into contigs and scaffolds
 for de-novo assembly of short-reads using de Bruijn graphs.\\ for de-novo assembly of short-reads using de Bruijn graphs.\\
 Can use a hierarchy of sizes of paired-end data.\\ Can use a hierarchy of sizes of paired-end data.\\
 Has been successfully used to sequence the Panda and Human genomes.\\ Has been successfully used to sequence the Panda and Human genomes.\\
-Quality seems good.  They ran Panda genome on 256GB workstation with 32 CPUs.+Quality seems good.  They ran Panda genome on 512GB workstation with 32 CPUs.
  
 There is a [[http://​soap.genomics.org.cn/​soapdenovo.html|description]] which contains a There is a [[http://​soap.genomics.org.cn/​soapdenovo.html|description]] which contains a
 download link. download link.
  
-Created by BGI - Beijing Genomics Institute ​[[http://​en.wikipedia.org/​wiki/​Beijing_Genomics_Institute|wiki]].+Created by BGI - [[wp>Beijing_Genomics_Institute]].
  
 **The sequence and de novo assembly of the giant panda genome** **The sequence and de novo assembly of the giant panda genome**
 [(cite:​panda>​ [(cite:​panda>​
-  ​Nature 463, 311-317 (21 January 2010)\\ +The sequence and de novo assembly of the giant panda genome 
-  doi:​[[10.1038/​nature08696|10.1038/​nature08696]];​ Received 19 August 2009; Accepted 24 November 2009; Published online 13 December 2009 +Nature 463, 311-317 (21 January 2010)\\ 
-  The sequence and de novo assembly of the giant panda genome +doi:[[http://​dx.doi.org/​10.1038/​nature08696|10.1038/​nature08696]];​\\ 
-[[http://​www.nature.com/​nature/​journal/​v463/​n7279/​full/​nature08696.html|article]]+Received 19 August 2009; Accepted 24 November 2009; Published online 13 December 2009 
 +)]
  
  
 Downloaded the binaries for SOAPdenovo and [[http://​soap.genomics.org.cn/​about.html#​resource2|GapCloser]]. I copied these binaries to the /bin folder since there are only two, and they at least correctly display a help message when you run them without arguments. Downloaded the binaries for SOAPdenovo and [[http://​soap.genomics.org.cn/​about.html#​resource2|GapCloser]]. I copied these binaries to the /bin folder since there are only two, and they at least correctly display a help message when you run them without arguments.
  
 +==== Method ====
 +The method for SOAPdenovo is described in the paper [[http://​genome.cshlp.org/​content/​20/​2/​265.full|"​De novo assembly of human genomes with massively parallel short read sequencing"​]] by Li et al.
 +
 +=== Short Read Data ===
 +Fragment and paired-end libraries are sequenced using various insert sizes. They used read lengths from 35 to 75 bp and insert sizes of 140 bp, 440 bp, 2.6 kb, 6 kb, and 9.6 kb.
 +
 +Basic error correction was performed on the reads using K-mer counting to reduce the memory usage when constructing the de Bruijn graph. This was done by identifying low frequency (occurring <3 times) 17-mers and correcting these K-mers to the candidate with the highest frequency. ​
 +
 +=== De Bruijn Graph === 
 +The next step is to build the de Bruijn graph to represent overlap of k-mers. 25-mers were used in their assembly.
 +Only the single-end and paired-end reads with short insert sizes (<1 kb) were used in the graph due the high probability of chimeric reads in the long-insert pairs from the circularizing and fragmentation process. Further error correction is done using the de Bruijn graph. ​
 +
 +    * Clip tips (low coverage paths that lead to dead ends)
 +    * Remove low coverage links
 +    * Resolve tiny repeats greater than K, but less than the read lengths.
 +    * Merge bubbles (paths with the same start and end). These can represent an error or a true polymorphism.
 +
 +==== Quirks ====
 +
 +=== NO FASTA, ONLY FASTQ ===
 +Unable to get SOAPdenovo to read any kind of FASTA file, despite
 +the documentation FASTA examples. ​ Tried many variants of the FASTA file,
 +even tried all 5 versions available for download, but
 +could not get it to work.  The other example shows the use of FASTQ.
 +Found and installed sff2fastq utility. Made FASTQ version of the 454 data.
 +Was able to get SOAPdenovo to run finally.
 +
 +Perhaps it just won't take the fasta input by itself.
 +It might work if you include a qual file with your fasta.
 +
 +==== Using SOAPdenovo ====
 +
 +SOAPdenovo has three executables each tuned for a different range of k-mer sizes (SOAPdenovo-31mer,​ SOAPdenovo-63mer,​ SOAPdenovo-127mer). For example, ''​SOAPdenovo-31mer''​ works best on k-mer sizes up to and including 31. For larger k-mers than 31 and lower than 64, use ''​SOAPdenovo-63mer''​.
 +
 +SOAPdenovo requires a configuration file that describes the libraries that will be used in the assembly. A library entry is required for each read file or pair of read files in the case of paired-end reads. Here is an example of the 5 library entries for 1 lane of run1.
 +
 +<​code>​
 +[LIB]
 +#average insert size
 +avg_ins=150
 +
 +#if sequence needs to be reversed ​
 +reverse_seq=0
 +
 +#in which part(s) the reads are used
 +asm_flags=3
 +
 +#in which order the reads are used while scaffolding
 +rank=1
 +
 +#fastq file for read 1
 +q1=/​campusdata/​BME235/​data/​slug/​clean/​run1_seqprep_quake/​s_1_1_qseq_seqprep.cor.fastq.gz
 +#fastq file for read 2 always follows fastq file for read 1
 +q2=/​campusdata/​BME235/​data/​slug/​clean/​run1_seqprep_quake/​s_1_2_qseq_seqprep.cor.fastq.gz
 +
 +[LIB]
 +reverse_seq=0
 +asm_flags=3
 +rank=1
 +q=/​campusdata/​BME235/​data/​slug/​clean/​run1_seqprep_quake/​s_1_1_qseq_seqprep.cor_single.fastq.gz
 +
 +[LIB]
 +reverse_seq=1
 +asm_flags=3
 +rank=1
 +q=/​campusdata/​BME235/​data/​slug/​clean/​run1_seqprep_quake/​s_1_2_qseq_seqprep.cor_single.fastq.gz
 +
 +[LIB]
 +reverse_seq=0
 +asm_flags=3
 +rank=1
 +q=/​campusdata/​BME235/​data/​slug/​clean/​run1_seqprep_quake/​s_1_merged_qseq_seqprep.cor.fastq.gz
 +
 +</​code>​
 +
 +
 +
 +==== Installing ====
 +
 +  cd /​campusdata/​BME235/​programs
 +  wget http://​soap.genomics.org.cn/​down/​SOAPdenovo-v1.04.tgz
 +  tar xfz SOAPdenovo-v1.04.tgz
 +  mv SOAPdenovo_Release1.04 SOAPdenovo
 +  mv SOAPdenovo-v1.04.tgz SOAPdenovo/
 +  cd SOAPdenovo
 +  cp SOAPdenovo ../../bin/
  
  
 ==== Website ==== ==== Website ====
-[[http://?]]+[[http://soap.genomics.org.cn/​soapdenovo.html]]
  
 ==== Source with Binaries and Documentation ==== ==== Source with Binaries and Documentation ====
-[[http://?]]+[[http://soap.genomics.org.cn/​down/​]]
  
 ===== References ===== ===== References =====
archive/bioinformatic_tools/soapdenovo.1271241884.txt.gz · Last modified: 2010/04/14 10:44 by galt