**(1) Mapping to the reference genome (STAR)** | **(1) Mapping to the reference genome (STAR)** | ||||
Aligning RNAseq data to a reference genome is complicates by RNA splicing, [GATK RNAseq variant calling best practice](<https://software.broadinstitute.org/gatk/documentation/article.php?id=3891>) recommend [STAR](<https://github.com/alexdobin/STAR>). | |||||
Aligning RNAseq data to a reference genome is complicates by RNA splicing, [GATK RNAseq variant calling best practice](<https://software.broadinstitute.org/gatk/documentation/article.php?id=3891>) recommends [STAR](<https://github.com/alexdobin/STAR>). | |||||
Generate genome index files: | Generate genome index files: | ||||
```bash | ```bash | ||||
java -jar GenomeAnalysisTK.jar -T VariantFiltration -R hg_19.fasta -V input.vcf -window 35 -cluster 3 -filterName FS -filter "FS > 30.0" -filterName QD -filter "QD < 2.0" -o output.vcf | java -jar GenomeAnalysisTK.jar -T VariantFiltration -R hg_19.fasta -V input.vcf -window 35 -cluster 3 -filterName FS -filter "FS > 30.0" -filterName QD -filter "QD < 2.0" -o output.vcf | ||||
``` | ``` | ||||
**Reference** | |||||
1. GATK best practice https://software.broadinstitute.org/gatk/documentation/article.php?id=3891 | |||||
2. Sentieon manual https://support.sentieon.com/manual/RNA_call/rna/ | |||||
3. STAR github https://github.com/alexdobin/STAR | |||||
4. Picard github https://github.com/broadinstitute/picard | |||||
5. Picard manual https://broadinstitute.github.io/picard/ | |||||
6. samtools manual http://www.htslib.org/doc/samtools.html |
"{{ project_name }}.fastq_2": "{{ read2 }}", | "{{ project_name }}.fastq_2": "{{ read2 }}", | ||||
"{{ project_name }}.dbmills_dir": "oss://pgx-reference-data/GRCh38.d1.vd1/", | "{{ project_name }}.dbmills_dir": "oss://pgx-reference-data/GRCh38.d1.vd1/", | ||||
"{{ project_name }}.cluster_config": "{{ cluster if cluster != '' else 'OnDemand ecs.sn1ne.8xlarge img-ubuntu-vpc' }}", | "{{ project_name }}.cluster_config": "{{ cluster if cluster != '' else 'OnDemand ecs.sn1ne.8xlarge img-ubuntu-vpc' }}", | ||||
"{{ project_name }}.SAref_dir": "File", | |||||
"{{ project_name }}.SAref_dir": "oss://chinese-quartet/quartet-storage-data/reference_data/STAR_GRCh38/", | |||||
"{{ project_name }}.SAMdocker": "registry-vpc.cn-shanghai.aliyuncs.com/pgx-docker-registry/samtools", | "{{ project_name }}.SAMdocker": "registry-vpc.cn-shanghai.aliyuncs.com/pgx-docker-registry/samtools", | ||||
"{{ project_name }}.STref_dir": "oss://chinese-quartet/quartet-storage-data/reference_data/STAR_GRCh38/", | |||||
"{{ project_name }}.STref_dir": "oss://chinese-quartet/quartet-storage-data/reference_data/", | |||||
"{{ project_name }}.db_mills": "Mills_and_1000G_gold_standard.indels.hg38.vcf", | "{{ project_name }}.db_mills": "Mills_and_1000G_gold_standard.indels.hg38.vcf", | ||||
"{{ project_name }}.sample": "{{ sample }}", | "{{ project_name }}.sample": "{{ sample }}", | ||||
"{{ project_name }}.dbsnp": "dbsnp_146.hg38.vcf", | "{{ project_name }}.dbsnp": "dbsnp_146.hg38.vcf", |