HOWTO

Building indexes

Depend on your purpose, you have to download reference sequence, gene annotation and SNP files.
We also provides scripts to build indexes. [Download Link]

Prepare data

  1. Download reference
    $ wget ftp://ftp.ensembl.org/pub/release-84/fasta/homo_sapiens/dna/Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz
    $ gzip -d Homo_sapiens.GRCh38.dna.primary_assembly.fa.gz
    $ mv Homo_sapiens.GRCh38.dna.primary_assembly.fa genome.fa
    
  2. Download GTF and make exon, splicesite file.
    If you want to build HFM index, you can skip this step.
    $ wget ftp://ftp.ensembl.org/pub/release-84/gtf/homo_sapiens/Homo_sapiens.GRCh38.84.gtf.gz  
    $ gzip -d Homo_sapiens.GRCh38.84.gtf.gz
    $ mv Homo_sapiens.GRCh38.84.gtf genome.gtf
    $ hisat2_extract_splice_sites.py genome.gtf > genome.ss
    $ hisat2_extract_exons.py genome.gtf > genome.exon
    
  3. Download SNP
    If you want to build HFM index, you can skip this step.
    $ wget http://hgdownload.cse.ucsc.edu/goldenPath/hg38/database/snp144Common.txt.gz
    $ gzip -d snp144Common.txt.gz
    

    Convert chromosome names of UCSC Database to Ensembl Annotation

    $ awk 'BEGIN{OFS="\t"} {if($2 ~ /^chr/) {$2 = substr($2, 4)}; if($2 == "M") {$2 = "MT"} print}' snp144Common.txt > snp144Common.txt.ensembl
    

    make SNPs and haplotype file

    $ hisat2_extract_snps_haplotypes_UCSC.py genome.fa snp144Common.txt.ensembl genome
    

Build HFM index

It takes about 20 minutes(depend on HW spec) to build index, and requires at least 6GB memory.

$ hisat2-build -p 16 genome.fa genome

Build HGFM index with SNPs

$ hisat2-build -p 16 --snp genome.snp --haplotype genome.haplotype genome.fa genome_snp

Build HGFM index with transcripts

It takes about 1 hour(depend on HW spec) to build index, and requires at least 160GB memory.

$ hisat2-build -p 16 --exon genome.exon --ss genome.ss genome.fa genome_tran

Build HGFM index with SNPs and transcripts

$ hisat2-build -p 16 --snp genome.snp --haplotype genome.haplotype --exon genome.exon --ss genome.ss genome.fa genome_snp_tran