Snakemake：snakemake 管道中的 MissingInputException
Snakemake: MissingInputException in snakemake pipeline
我正在尝试 SnakeMake 管道，但遇到一个我真的不明白的错误。
我有一个目录 (raw_data)，其中有输入文件：
ll /home/nico/labo/etudes/Optimal/data/raw_data
total 41M
drwxrwxr-x 2 nico nico 4,0K mars 6 16:09 ./
drwxrwxr-x 11 nico nico 4,0K mars 6 16:14 ../
-rw-rw-r-- 1 nico nico 15M févr. 27 12:21 sampleA_R1.fastq.gz
-rw-rw-r-- 1 nico nico 19M févr. 27 12:22 sampleA_R2.fastq.gz
-rw-rw-r-- 1 nico nico 3,4M févr. 27 12:21 sampleB_R1.fastq.gz
-rw-rw-r-- 1 nico nico 4,3M févr. 27 12:22 sampleB_R2.fastq.gz
此目录包含 2 个示例的 4 个文件。
我为名为 config_snakemake_Optimal_mapping_BaL.json:
的 SnakeMake 管道创建了一个配置 json 文件
{
"fastqExtension": "fastq.gz",
"fastqDir": "/home/nico/labo/etudes/Optimal/data/raw_data",
"outputDir": "/home/nico/labo/etudes/Optimal/data/mapping_BaL",
"logDir": "logs",
"reference": {
"fasta": "/home/nico/labo/references/genomes/HIV1/BaL_AY713409/BaL_AY713409.fasta",
"index": "/home/nico/labo/references/genomes/HIV1/BaL_AY713409/BaL_AY713409.fasta.bwt"
}
}
最后是 SnakeMake 文件 snakefile_bwa_samtools.py:
import subprocess
from os.path import join
### Globals ---------------------------------------------------------------------
# A Snakemake regular expression matching fastq files.
SAMPLES, = glob_wildcards(join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]))
print(SAMPLES)
### Rules -----------------------------------------------------------------------
# Pipeline output files
rule all:
input: expand(join(config["outputDir"], "{sample}.bam.bai"), sample=SAMPLES)
# Reads alignment on reference genome and BAM file creation
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1_ID = "{sample}_R1."+config["fastqExtension"],
fq2_ID = "{sample}_R2."+config["fastqExtension"],
fq1 = join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]),
fq2 = join(config["fastqDir"], "{sample}_R2."+config["fastqExtension"])
output:
temp(join(config["outputDir"], "{sample}.bamUnsorted"))
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {input.fq1_ID} and {input.fq2_ID} on {input.fasta} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
# Sorting the BAM files on genomic positions
rule bam_sort:
input:
join(config["outputDir"], "{sample}.bamUnsorted")
output:
join(config["outputDir"], "{sample}.bam")
log:
join(config["outputDir"], config["logDir"], "{sample}.samtools_sort.log")
version:
subprocess.getoutput(
"samtools --version | "
"head -1 | "
"cut -d' ' -f2"
)
message:
"Genomic sorting of {input} with samtools version {version}."
shell:
"samtools sort -f {input} {output} 2> {log}"
# Indexing the BAM files
rule bam_index:
input:
join(config["outputDir"], "{sample}.bam")
output:
join(config["outputDir"], "{sample}.bam.bai")
message:
"Indexing {input}."
shell:
"samtools index {input}"
我运行这条管道：
snakemake --cores 3 --snakefile /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py --configfile /home/nico/labo/etudes/Optimal/data/snakemake_config_files/config_snakemake_Optimal_mapping_BaL.json
我得到以下错误输出：
['sampleB', 'sampleA']
MissingInputException in line 18 of /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py:
Missing input files for rule bwa_mem_to_bam:
sampleB_R1.fastq.gz
sampleB_R2.fastq.gz
或取决于时刻：
['sampleB', 'sampleA']
PeriodicWildcardError in line 40 of /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py:
The value _unsorted in wildcard sample is periodically repeated (sampleB_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted). This would lead to an infinite recursion. To avoid this, e.g. restrict the wildcards in this rule to certain values.
样本在列表中出现时被正确检测到（输出类型的第一行），我肯定在乱用规则 bwa_mem_to_bam 中的通配符，但我真的不明白为什么..
有什么线索吗？
最后，我成功地删除了 rule bwa_mem_to_bam 中的 fq1_ID 和 fq2_ID 变量，并替换了规则 input.fq1_ID 的 message和 input.fq2_ID 通过 input.fq1 和 input.fq2.
消息不够优雅，但管道 运行 正确。还是不明白哪里错了，如果有人能解释一下，我还在听！
rule bwa_mem_to_bam的正确代码：
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1 = join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]),
fq2 = join(config["fastqDir"], "{sample}_R2."+config["fastqExtension"])
output:
temp(join(config["outputDir"], "{sample}.bamUnsorted"))
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {input.fq1} and {input.fq2} on {input.fasta} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
我很快看了你的代码。
为什么第一个没有成功？
当你声明 fq1_ID 和 fq1 时，与示例 2 相同。你没有分配相同的字符串。对于 fq1，您为文件添加一个目录 对于 fq1_ID 不存在，因此 snakemake 正在工作目录中搜索它（如果未设置 -d 选项，则为当前目录）a文件名与您的字符串。因为这些变量在输入部分。
所以通过删除两个 fq1/2_ID，它将消除所有文件搜索问题。
雨果
感谢雨果检查我的代码和你的解释，很有道理！
今天早上醒来我终于有了一个闪现的想法（最好的），并意识到我忽略了规则的 params 部分，fq1_ID 和 fq2_ID 不是输入而是参数..
我将代码更改为：
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1 = join(config["fastqDir"], "{sample}_R1.fastq.gz"),
fq2 = join(config["fastqDir"], "{sample}_R2.fastq.gz")
output:
temp(join(config["outputDir"],"{sample}_unsorted.bam"))
params:
fq1_ID = "{sample}_R1.fastq.gz",
fq2_ID = "{sample}_R2.fastq.gz",
ref_ID = os.path.basename(config["reference"]["fasta"])
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {params.fq1_ID} and {params.fq2_ID} on {params.ref_ID} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
而且效果很好！
snakemake --cores 3 --snakefile /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py --configfile /home/nico/labo/etudes/Optimal/data/snakemake_config_files/config_snakemake_Optimal_mapping_BaL.json
Provided cores: 3
Rules claiming more threads will be scaled down.
Job counts:
count jobs
1 all
2 bam_index
2 bam_sort
2 bwa_mem_to_bam
7
Alignment of sampleB_R1.fastq.gz and sampleB_R2.fastq.gz on BaL_AY713409.fasta with BWA version 0.7.12.
Alignment of sampleA_R1.fastq.gz and sampleA_R2.fastq.gz on BaL_AY713409.fasta with BWA version 0.7.12.
1 of 7 steps (14%) done
Genomic sorting of sampleB_unsorted.bam with samtools version 1.2.
Removing temporary output file /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleB_unsorted.bam.
2 of 7 steps (29%) done
Indexing sampleB.bam.
3 of 7 steps (43%) done
4 of 7 steps (57%) done
Genomic sorting of sampleA_unsorted.bam with samtools version 1.2.
Removing temporary output file /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleA_unsorted.bam.
5 of 7 steps (71%) done
Indexing sampleA.bam.
6 of 7 steps (86%) done
localrule all:
input: /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleB.bam.bai, /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleA.bam.bai
7 of 7 steps (100%) done
终于收到我的正确消息：
- sampleB_R1.fastq.gz 和 sampleB_R2.fastq.gz 对齐
BaL_AY713409.fasta BWA 版本 0.7.12。
- sampleA_R1.fastq.gz 和 sampleA_R2.fastq.gz 在 BaL_AY713409.fasta 上的对齐
BWA 版本 0.7.12.
我正在尝试 SnakeMake 管道，但遇到一个我真的不明白的错误。
我有一个目录 (raw_data)，其中有输入文件：
ll /home/nico/labo/etudes/Optimal/data/raw_data
total 41M
drwxrwxr-x 2 nico nico 4,0K mars 6 16:09 ./
drwxrwxr-x 11 nico nico 4,0K mars 6 16:14 ../
-rw-rw-r-- 1 nico nico 15M févr. 27 12:21 sampleA_R1.fastq.gz
-rw-rw-r-- 1 nico nico 19M févr. 27 12:22 sampleA_R2.fastq.gz
-rw-rw-r-- 1 nico nico 3,4M févr. 27 12:21 sampleB_R1.fastq.gz
-rw-rw-r-- 1 nico nico 4,3M févr. 27 12:22 sampleB_R2.fastq.gz
此目录包含 2 个示例的 4 个文件。
我为名为 config_snakemake_Optimal_mapping_BaL.json:
{
"fastqExtension": "fastq.gz",
"fastqDir": "/home/nico/labo/etudes/Optimal/data/raw_data",
"outputDir": "/home/nico/labo/etudes/Optimal/data/mapping_BaL",
"logDir": "logs",
"reference": {
"fasta": "/home/nico/labo/references/genomes/HIV1/BaL_AY713409/BaL_AY713409.fasta",
"index": "/home/nico/labo/references/genomes/HIV1/BaL_AY713409/BaL_AY713409.fasta.bwt"
}
}
最后是 SnakeMake 文件 snakefile_bwa_samtools.py:
import subprocess
from os.path import join
### Globals ---------------------------------------------------------------------
# A Snakemake regular expression matching fastq files.
SAMPLES, = glob_wildcards(join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]))
print(SAMPLES)
### Rules -----------------------------------------------------------------------
# Pipeline output files
rule all:
input: expand(join(config["outputDir"], "{sample}.bam.bai"), sample=SAMPLES)
# Reads alignment on reference genome and BAM file creation
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1_ID = "{sample}_R1."+config["fastqExtension"],
fq2_ID = "{sample}_R2."+config["fastqExtension"],
fq1 = join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]),
fq2 = join(config["fastqDir"], "{sample}_R2."+config["fastqExtension"])
output:
temp(join(config["outputDir"], "{sample}.bamUnsorted"))
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {input.fq1_ID} and {input.fq2_ID} on {input.fasta} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
# Sorting the BAM files on genomic positions
rule bam_sort:
input:
join(config["outputDir"], "{sample}.bamUnsorted")
output:
join(config["outputDir"], "{sample}.bam")
log:
join(config["outputDir"], config["logDir"], "{sample}.samtools_sort.log")
version:
subprocess.getoutput(
"samtools --version | "
"head -1 | "
"cut -d' ' -f2"
)
message:
"Genomic sorting of {input} with samtools version {version}."
shell:
"samtools sort -f {input} {output} 2> {log}"
# Indexing the BAM files
rule bam_index:
input:
join(config["outputDir"], "{sample}.bam")
output:
join(config["outputDir"], "{sample}.bam.bai")
message:
"Indexing {input}."
shell:
"samtools index {input}"
我运行这条管道：
snakemake --cores 3 --snakefile /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py --configfile /home/nico/labo/etudes/Optimal/data/snakemake_config_files/config_snakemake_Optimal_mapping_BaL.json
我得到以下错误输出：
['sampleB', 'sampleA']
MissingInputException in line 18 of /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py:
Missing input files for rule bwa_mem_to_bam:
sampleB_R1.fastq.gz
sampleB_R2.fastq.gz
或取决于时刻：
['sampleB', 'sampleA']
PeriodicWildcardError in line 40 of /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py:
The value _unsorted in wildcard sample is periodically repeated (sampleB_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted_unsorted). This would lead to an infinite recursion. To avoid this, e.g. restrict the wildcards in this rule to certain values.
样本在列表中出现时被正确检测到（输出类型的第一行），我肯定在乱用规则 bwa_mem_to_bam 中的通配符，但我真的不明白为什么..
有什么线索吗？
最后，我成功地删除了 rule bwa_mem_to_bam 中的 fq1_ID 和 fq2_ID 变量，并替换了规则 input.fq1_ID 的 message和 input.fq2_ID 通过 input.fq1 和 input.fq2.
消息不够优雅，但管道 运行 正确。还是不明白哪里错了，如果有人能解释一下，我还在听！
rule bwa_mem_to_bam的正确代码：
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1 = join(config["fastqDir"], "{sample}_R1."+config["fastqExtension"]),
fq2 = join(config["fastqDir"], "{sample}_R2."+config["fastqExtension"])
output:
temp(join(config["outputDir"], "{sample}.bamUnsorted"))
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {input.fq1} and {input.fq2} on {input.fasta} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
我很快看了你的代码。
为什么第一个没有成功？
当你声明 fq1_ID 和 fq1 时，与示例 2 相同。你没有分配相同的字符串。对于 fq1，您为文件添加一个目录 对于 fq1_ID 不存在，因此 snakemake 正在工作目录中搜索它（如果未设置 -d 选项，则为当前目录）a文件名与您的字符串。因为这些变量在输入部分。
所以通过删除两个 fq1/2_ID，它将消除所有文件搜索问题。
雨果
感谢雨果检查我的代码和你的解释，很有道理！
今天早上醒来我终于有了一个闪现的想法（最好的），并意识到我忽略了规则的 params 部分，fq1_ID 和 fq2_ID 不是输入而是参数..
我将代码更改为：
rule bwa_mem_to_bam:
input:
index = config["reference"]["index"],
fasta = config["reference"]["fasta"],
fq1 = join(config["fastqDir"], "{sample}_R1.fastq.gz"),
fq2 = join(config["fastqDir"], "{sample}_R2.fastq.gz")
output:
temp(join(config["outputDir"],"{sample}_unsorted.bam"))
params:
fq1_ID = "{sample}_R1.fastq.gz",
fq2_ID = "{sample}_R2.fastq.gz",
ref_ID = os.path.basename(config["reference"]["fasta"])
version:
subprocess.getoutput(
"man bwa | tail -n 1 | cut -d ' ' -f 1 | cut -d '-' -f 2"
)
log:
join(config["outputDir"], config["logDir"], "{sample}.bwa_mem.log")
message:
"Alignment of {params.fq1_ID} and {params.fq2_ID} on {params.ref_ID} with BWA version {version}."
shell:
"bwa mem {input.fasta} {input.fq1} {input.fq2} 2> {log} | samtools view -Sbh - > {output}"
而且效果很好！
snakemake --cores 3 --snakefile /home/nico/labo/scripts/pipeline_illumina/snakefile_bwa_samtools.py --configfile /home/nico/labo/etudes/Optimal/data/snakemake_config_files/config_snakemake_Optimal_mapping_BaL.json
Provided cores: 3
Rules claiming more threads will be scaled down.
Job counts:
count jobs
1 all
2 bam_index
2 bam_sort
2 bwa_mem_to_bam
7
Alignment of sampleB_R1.fastq.gz and sampleB_R2.fastq.gz on BaL_AY713409.fasta with BWA version 0.7.12.
Alignment of sampleA_R1.fastq.gz and sampleA_R2.fastq.gz on BaL_AY713409.fasta with BWA version 0.7.12.
1 of 7 steps (14%) done
Genomic sorting of sampleB_unsorted.bam with samtools version 1.2.
Removing temporary output file /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleB_unsorted.bam.
2 of 7 steps (29%) done
Indexing sampleB.bam.
3 of 7 steps (43%) done
4 of 7 steps (57%) done
Genomic sorting of sampleA_unsorted.bam with samtools version 1.2.
Removing temporary output file /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleA_unsorted.bam.
5 of 7 steps (71%) done
Indexing sampleA.bam.
6 of 7 steps (86%) done
localrule all:
input: /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleB.bam.bai, /home/nico/labo/etudes/Optimal/data/mapping_BaL/sampleA.bam.bai
7 of 7 steps (100%) done
终于收到我的正确消息：
- sampleB_R1.fastq.gz 和 sampleB_R2.fastq.gz 对齐 BaL_AY713409.fasta BWA 版本 0.7.12。
- sampleA_R1.fastq.gz 和 sampleA_R2.fastq.gz 在 BaL_AY713409.fasta 上的对齐 BWA 版本 0.7.12.