跳转到内容

组装质量评估

作用:用回贴深度、BUSCO 与 Merqury 三块独立评估最终组装的完整度与碱基准确性。 入口:01_mapping/{01_ont,02_hifi,03_wgs,04_pan_wgs}/、02_busco/、03_merqury/{01_hifi,02_wgs,03_pan_wgs}/ 下的脚本 输入:组装 ``{PROJ_LOTUS_ZC}/Gifu/00.assemble_result/Gifu_v1.0.fasta;reads ONT Gifu-jing-ye.pass.ul.fa、HiFi Gifu_hifi.fastq.gz、WGS Gifu_1-R{1,2}.fastq.gz;窗口 Gifu_{100k,10k}.bed;HiFi k-mer 库 ccs.merylDB;BUSCO lineage embryophyta_odb10。 输出:gifu_{ont,hifi,wgs}_{100k,10k}_coverage.txt;BUSCO Gifu_lotus/;Merqury Gifu_hifimer、Gifu_wgsmer、Gifu_<sample>_wgsmer;pan-WGS {Gifu,MG20}_Clean_R{1,2}.fq 与 MG20_subsample_{summary,scientific_name}.txt。 运行:进入相应子目录后 bash <script> 工具:minimap2 -ax map-ont/-ax map-pb、bwa mem;samtools view -b -q 30、sort、flagstat;bedtools coverage -mean(100 kb 与 10 kb 窗口);BBMap filterbyname.sh;BLAST+ blastn -outfmt 5;BUSCO -m genome --offline --cpu 64;Merqury 与 meryl count k=21。 注意:ont.sh、ccs.sh、bwa.sh 与 04_pan_wgs/samtools.sh 的比对命令均被注释,脚本只对现成的 ont_q30.bam、ccs_q30.bam、wgs_q30.bam 与 *_unmapped.literal.ids 重跑下游覆盖度与 reads 过滤,须先自备这些文件。

English **Purpose**: Assess the finished assembly in three independent blocks: read-remapping depth, BUSCO gene-space completeness and Merqury k-mer QV. **Entry point**: scripts under `01_mapping/{01_ont,02_hifi,03_wgs,04_pan_wgs}/`, `02_busco/` and `03_merqury/{01_hifi,02_wgs,03_pan_wgs}/` **Inputs**: assembly ``{PROJ_LOTUS_ZC}`/Gifu/00.assemble_result/Gifu_v1.0.fasta`; reads ONT `Gifu-jing-ye.pass.ul.fa`, HiFi `Gifu_hifi.fastq.gz`, WGS `Gifu_1-R{1,2}.fastq.gz`; windows `Gifu_{100k,10k}.bed`; HiFi k-mer database `ccs.merylDB`; BUSCO lineage `embryophyta_odb10`. **Outputs**: `gifu_{ont,hifi,wgs}_{100k,10k}_coverage.txt`; BUSCO `Gifu_lotus/`; Merqury `Gifu_hifimer`, `Gifu_wgsmer`, `Gifu__wgsmer`; pan-WGS `{Gifu,MG20}_Clean_R{1,2}.fq` and `MG20_subsample_{summary,scientific_name}.txt`. **Run**: `cd` into the relevant sub-directory and run `bash