offtracker 2.7.8__zip → 2.10.0__zip

{offtracker-2.7.8 → offtracker-2.10.0}/offtracker/X_sequence.py

@@ -3,6 +3,7 @@ import math
 import pandas as pd
 from itertools import product
 import numpy as np
+import os, glob
 
 ambiguous_nt = {'A': ['A'],
                 'T': ['T'],
@@ -19,7 +20,7 @@ ambiguous_nt = {'A': ['A'],
                 'H': ['A', 'C', 'T'],
                 'D': ['A', 'G', 'T'],
                 'B': ['C', 'G', 'T'],
-                'N': ['A', 'T', 'C', 'G']}
+                'N': ['A', 'C', 'G', 'T']}
 
 def is_seq_valid(sequence, extra=True, ambiguous_nt=ambiguous_nt):
     if extra:
@@ -43,12 +44,24 @@ def possible_seq(sequence):
         raise KeyError(f'Unvalid character \'{valid_check}\' in sequence')
     return sequences
 
+# 包含 degenerate base pairs
+def get_base_score(base1, base2, exact_score=2, partial_match=2, mismatch_score=0.01):
+    base1 = ambiguous_nt[base1]
+    base2 = ambiguous_nt[base2]
+    if base1 == base2:
+        return exact_score
+    if list(np.union1d(base1,base2)) == base1 or list(np.union1d(base1,base2)) == base2:
+        # 其中一个是子集，注意顺序不一致会导致不等，所以必须排好序
+        return partial_match
+    return mismatch_score
+
+
 def complement(seq):
-    complement = {'A': 'T', 'C': 'G', 'G': 'C', 'T': 'A', 'N': 'N', '-':'-',
+    dict_complement = {'A': 'T', 'C': 'G', 'G': 'C', 'T': 'A', 'N': 'N', '-':'-',
                   'M': 'K', 'R': 'Y', 'W': 'W', 'S': 'S', 'Y': 'R', 'K':'M',
                   'V': 'B', 'H': 'D', 'D': 'H', 'B': 'V'} 
     bases = list(seq) 
-    letters = [complement[base] for base in bases] 
+    letters = [dict_complement[base] for base in bases] 
     return ''.join(letters)
 
 def reverse(seq):
@@ -100,14 +113,107 @@ def add_ID(df, chr_col=0, midpoint='cleavage_site'):#, midpoint='midpoint'):
 	df.loc[point_tail>=500,'ID_2'] = df[chr_col_name] + ':' + (point_head+1).astype(str)
 	return df
 
+
+
+def detect_fastq(folder, n_subfolder, NGS_type='paired-end'):
+    """
+    搜索 folder 的 n级子目录下的所有 fastq/fastq.gz/fq/fq.gz 文件
+    paired-end 模式 : 识别 2.fq/2.fastq 为 paired-end 的 R2 文件，并验证对应 R1 文件
+    single-end 模式 : 所有 fastq/fastq.gz/fq/fq.gz 文件都视为 single-end 文件
+    
+    不建议 2. 和 fq/fastq 之间有其他字符，如 2.trimmed.fq.gz，因为中间字符不确定，使用通配符容易误判文件名其他的2.
+    样本名不要带点，建议用_分割特征，同特征内分割不要用_可以用-，如 sample_day-hour_type_batch_rep_1.fq.gz
+
+    Input
+    ----------
+    folder : 根目录
+    n_subfolder : n级子目录
+
+    Parameter
+    ----------
+    NGS_type : 'paired-end' or 'single-end'
+    
+    Output
+    ----------
+    sample_names : 识别的样品名
+    files_R1 : R1文件的完整路径
+    files_R2 : R2文件的完整路径
+
+    """
+    # import os, sys, glob
+    # import pandas as pd
+    if NGS_type == 'paired-end':
+        print('paired-end mode')
+        files_R2 = []
+        # 支持四种文件扩展名
+        # 个人习惯包含绝对路径
+        for fastq in ['*2.fq','*2.fastq','*2.fq.gz','*2.fastq.gz']:
+            fq_files = glob.glob( os.path.join(folder, n_subfolder*'*/', fastq ) )
+            print(f'{len(fq_files)} {fastq[2:]} samples detected')
+            files_R2.extend( fq_files )
+        #
+        if len(files_R2) > 0:
+            files_R2 = pd.Series(files_R2).sort_values().reset_index(drop=True)
+            # 拆分文件名
+            suffix = files_R2.str.extract('(\.fastq.*|\.fq.*)',expand=False)
+            prefix = files_R2.str.extract('(.*)(?:.fq|.fastq)',expand=False)
+            # 将 prefix 进一步拆分为 sample_dir （真样品名） 和 nametype （某种统一后缀），支持五种样本名后缀
+            nametype = []
+            sample_dir = []
+            for a_prefix in prefix:
+                for a_type in ['_trimmed_2', '_2_val_2','_R2_val_2','_R2','_2']:
+                    len_type = len(a_type)
+                    if a_prefix[-len_type:] == a_type:
+                        nametype.append(a_type)
+                        sample_dir.append(a_prefix[:-len_type])
+                        break
+            assert len(nametype) == len(files_R2), 'The file name pattern is invaild!'
+            nametype = pd.Series(nametype)
+            sample_dir = pd.Series(sample_dir)
+            # 根据 R2 文件，检查 R1 文件是否存在
+            files_R1 = sample_dir + nametype.str.replace('2','1') + suffix
+            for i in range(len(files_R1)):
+                assert os.path.exists(files_R1[i]), f'{files_R1[i]} not found!'
+            sample_names = sample_dir.apply(os.path.basename)
+        else:
+            print('No paired-end samples detected!')
+            sample_names = 'no sample'
+            files_R1 = []
+
+    elif NGS_type == 'single-end':
+        print('single-end mode')
+        files_R1 = []
+        files_R2 = [] # 占位
+        # 支持四种文件扩展名
+        # 个人习惯包含绝对路径
+        for fastq in ['*.fq','*.fastq','*.fq.gz','*.fastq.gz']:
+            fq_files = glob.glob( os.path.join(folder, n_subfolder*'*/', fastq ) )
+            print(f'{len(fq_files)} {fastq[1:]} samples detected')
+            files_R1.extend( fq_files )
+        files_R1 = pd.Series(files_R1).sort_values()
+        #
+        if len(files_R1) > 0:
+            # 拆分文件名
+            suffix = files_R1.str.extract('(\.fastq.*|\.fq.*)',expand=False)
+            prefix = files_R1.str.extract('(.*)(?:.fq|.fastq)',expand=False)
+            # 单端模式下，所有前缀都视为样品名
+            sample_names = prefix.apply(os.path.basename)
+        else:
+            print('No single-end samples detected!')
+            sample_names = 'no sample'
+            files_R1 = []
+
+    return sample_names, files_R1, files_R2
+
+
 def sgRNA_alignment(a_key, sgRNA, seq, frag_len, DNA_matrix=None, mismatch_score = 0.01, return_align=False):
     from Bio import pairwise2
     import numpy as np
     if DNA_matrix is None:
-        DNA_matrix = {('A','A'): 2, ('A','T'):0.01, ('A','C'):0.01, ('A','G'):0.01, ('A','N'):0.01,
-                    ('T','T'): 2, ('T','A'):0.01, ('T','C'):0.01, ('T','G'):0.01, ('T','N'):0.01,
-                    ('G','G'): 2, ('G','A'):0.01, ('G','C'):0.01, ('G','T'):0.01, ('G','N'):0.01,
-                    ('C','C'): 2, ('C','A'):0.01, ('C','G'):0.01, ('C','T'):0.01, ('C','N'):0.01,
+        DNA_matrix = {('A','A'): 2, ('A','T'):0.01, ('A','C'):0.01, ('A','G'):0.01, ('A','N'):2,
+                    ('T','T'): 2, ('T','A'):0.01, ('T','C'):0.01, ('T','G'):0.01, ('T','N'):2,
+                    ('G','G'): 2, ('G','A'):0.01, ('G','C'):0.01, ('G','T'):0.01, ('G','N'):2,
+                    ('C','C'): 2, ('C','A'):0.01, ('C','G'):0.01, ('C','T'):0.01, ('C','N'):2,
                     ('N','N'): 2, ('N','C'):2, ('N','A'): 2, ('N','G'): 2, ('N','T'): 2}        
     # a_key 是 pybedtools 得到的位置 chrA:X-Y 而 X 数字会往左多1bp
     alignments = pairwise2.align.localds( sgRNA, seq, DNA_matrix, -2, -2, penalize_extend_when_opening=False)

offtracker-2.10.0/offtracker/_version.py

@@ -0,0 +1,36 @@
+__version__ = "2.10.0"
+# 2023.08.11. v1.1.0	adding a option for not normalizing the bw file
+# 2023.10.26. v1.9.0	prerelease for v2.0
+# 2023.10.27. v2.0.0	大更新，还没微调
+# 2023.10.28. v2.1.0	修复bug，增加计算信号长度的功能
+# 2023.10.28. v2.2.0	修复bug，改变计算信号长度的算法
+# 2023.10.29. v2.3.0	增加 overall signal 计算
+# 2023.11.01. v2.3.1	增加 signal_only 选项
+# 2023.11.02. v2.3.2	修改 sample signal 和 group mean 的计算顺序
+# 2023.11.04. v2.3.3	修复 overall score 标准化时排序错误的问题
+# 2023.11.05. v2.3.4	修复判断单边溢出信号时的列名选取错误
+# 2023.11.13. v2.3.5	微调 track score
+# 2023.12.05. v2.3.6	candidates 增加 cleavage site，修正 alignment 有 deletion 会错位的 bug
+# 2023.12.05. v2.3.7	用 cleavage site 代替 midpoint # 还没改完
+# 2023.12.07. v2.3.8	df_score 增加 df_exp, df_ctr 各自列。修复没 df_ctr 时的 bug。track score 用 proximal
+# 2023.12.09. v2.4.0	为了兼顾 proximal 和 overall，当 normalized overall signal 高于 2 时，增加 overall signal 的加分
+# 2023.12.09. v2.5.0	尝试新的加权位置
+# 2023.12.10. v2.6.0	加入 trackseq v4 的计算分支，即考虑 Region 内的 positive_pct，避免短而尖锐的信号
+# 2023.12.10. v2.6.1	有些非特异信号数值很大，如果在 control 组是大负数，可能导致减 control 后假高信号，因此给负数一个 clip
+# 2023.12.30. v2.7.0	增加 X_offplot 模块，用于绘图
+# 2023.12.31. v2.7.1	control 的负数值 clip 由 -5 改为 -1，进一步减少假阳性。另外不加 overall 了
+# 2024.01.01. v2.7.2	权重改为 proximal + pct = 1 + 1. 防信号外溢假阳性标准由<0改为<=0
+# 2024.01.02. v2.7.3	flank regions 默认值改为 1000 2000 3000 5000。之前 control 的负数值 clip 相当于直接在 final score，现在改为每个单独 clip 后重新算 score，默认值为 CtrClip=-0.5
+# 2024.01.03. v2.7.4	更新了 blacklist.bed
+# 2024.01.04. v2.7.5	更新了 hg38 blacklist.bed
+# 2024.01.12. v2.7.6	修复小bug，输出 fdr 改为 <0.05。
+# 2024.01.23. v2.7.7	Snakefile_offtracker: add --fixedStep to bigwigCompare for not merging neighbouring bins with equal values.
+# 2024.02.01. v2.7.8	逐步添加 X_offplot.py 功能
+# 2024.06.02. v2.7.9	添加 offtracker_plot.py
+# 2024.06.03. v2.7.10	修复 bugs，offtable 添加 threshold = 2 的分界
+# 2024.06.04. v2.7.11	readme 修改
+# 2024.11.19. v2.7.12	offtracker_candidates.py 新增 --pam_location 参数指定 upstream 或 downstream，用于非 Cas9 情况
+# 2025.04.25. v2.8.0	修复了 offtracker candidates 会把小写序列转换成 N 的 bug
+# 2025.05.22. v2.9.0	翻新部分代码结构
+# 2025.06.05. v2.10.0	增加了QC模块。保留了负数score的记录，并在plot时显示为红字。增加了 "--ignore_chr" 用于跳过common chr过滤。
+

offtracker-2.10.0/offtracker/snakefile/Snakefile_QC.smk

@@ -0,0 +1,66 @@
+# 更新记录:
+# 2022.05.04. v1.0:    初步运行, fastp + multiqc
+# 2024.01.17. v2.0:    翻新结构，匹配 X_NGS 框架
+
+# 参数列表
+configfile: "config.yaml"
+
+### config['files_R1'], config['files_R2'] 为 dict型
+
+# # fastq 信息
+_files_R1 = config['files_R1'] # dict型, key 为 sample
+_files_R2 = config['files_R2'] # dict型, key 为 sample
+# # 输入输出文件夹
+# config['input_dir']
+_output_dir = config["output_dir"]
+# # 运行参数
+_thread = config['thread']
+# config['utility_dir']
+
+import os
+
+############################
+# conditional output_files #
+############################
+output_HT = expand( os.path.join(_output_dir,"{sample}_fastp.html"), sample=_files_R1)
+output_JS = expand( os.path.join(_output_dir,"{sample}_fastp.json"), sample=_files_R1)
+output_MQC = os.path.join(_output_dir,"MultiQC_Report_Raw.html")
+output_R1 = expand( os.path.join(_output_dir,"{sample}_trimmed_1.fq.gz"), sample=_files_R1) # dict 会自动迭代 keys
+output_R2 = expand( os.path.join(_output_dir,"{sample}_trimmed_2.fq.gz"), sample=_files_R1)
+
+output_files = output_HT + output_JS + [output_MQC] + output_R1 + output_R2
+
+rule all:
+    input:
+        output_files
+
+#######################
+## fastp and multiQC ##
+#######################
+rule QCtrim:
+    input:
+        R1=lambda w: _files_R1[w.sample],
+        R2=lambda w: _files_R2[w.sample]
+    threads:
+        _thread
+    output:
+        R1=os.path.join(_output_dir,"{sample}_trimmed_1.fq.gz"),
+        R2=os.path.join(_output_dir,"{sample}_trimmed_2.fq.gz"),
+        HT=os.path.join(_output_dir,"{sample}_fastp.html"),
+        JS=os.path.join(_output_dir,"{sample}_fastp.json")
+    shell:
+        """
+        fastp -i {input.R1} -I {input.R2} -o {output.R1} -O {output.R2} \
+        -h {wildcards.sample}_fastp.html -j {wildcards.sample}_fastp.json \
+        --length_required 10 --thread {threads} --detect_adapter_for_pe --disable_quality_filtering
+        """
+
+rule multiqc:
+    input:
+        expand( os.path.join(_output_dir,"{sample}_fastp.html"), sample=_files_R1 )
+    threads:
+        _thread
+    output:
+        os.path.join(_output_dir,"MultiQC_Report_Raw.html")
+    shell:
+        "multiqc {_output_dir} -n MultiQC_Report_Raw --outdir {_output_dir}"

offtracker-2.10.0/offtracker/snakefile/Snakefile_offtracker.smk

@@ -0,0 +1,249 @@
+# 2023.08.11. adding a option for not normalizing the bw file
+# 2024.01.23. add --fixedStep to bigwigCompare for not merging neighbouring bins with equal values.
+# 2025.05.22. refine the structure
+
+configfile: "config.yaml"
+
+# # fastq 信息
+_files_R1 = config['files_R1'] # dict型, key 为 sample
+_files_R2 = config['files_R2'] # dict型, key 为 sample
+# # 运行参数
+_output_dir = config["output_dir"]
+_thread = config['thread']
+_BinSize = str(config["binsize"])
+_normalize = config["normalize"]
+
+
+import os
+
+if _normalize == "True":
+    rule all:
+        input:
+            expand( os.path.join(_output_dir,"{sample}.fw.bed"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.rv.bed"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.fw.scaled.bw"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.rv.scaled.bw"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}." + _BinSize + ".add.bdg"),sample=_files_R1 ),
+elif _normalize == "False":
+    rule all:
+        input:
+            expand( os.path.join(_output_dir,"{sample}.fw.bed"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.rv.bed"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.fw.raw.bw"), sample=_files_R1 ),
+            expand( os.path.join(_output_dir,"{sample}.rv.raw.bw"), sample=_files_R1 ),
+else:
+    raise ValueError('Please provide "True" or "False" for "--normalize" when running offtracker_config.py')
+
+
+rule chromap:
+    input:
+        R1=lambda w: _files_R1[w.sample],
+        R2=lambda w: _files_R2[w.sample]
+    threads:
+        _threads
+    params:
+        index=config["index"],
+        fasta=config["fasta"]
+    output:
+        temp(os.path.join(_output_dir,"{sample}.chromapx.bed"))
+    shell:
+        """
+        chromap -l 3000 --low-mem --BED --remove-pcr-duplicates \
+        --min-read-length 10 --allocate-multi-mappings \
+        -x {params.index} -r {params.fasta} -t {threads} -1 {input.R1} -2 {input.R2} -o {output}
+        """
+
+if config["blacklist"] != 'none':
+    rule remove_blacklist:
+        input:
+            os.path.join(_output_dir,"{sample}.chromapx.bed")
+        threads:
+            _threads
+        params:
+            blacklist=config["blacklist"]
+        output:
+            temp(os.path.join(_output_dir,"{sample}.filtered.bed"))
+        shell:
+            "bedtools intersect -a {input} -b {params.blacklist} -v > {output}"
+
+    rule bed2fr:
+        input:
+            os.path.join(_output_dir,"{sample}.filtered.bed")
+        threads:
+            _threads
+        params:
+            dir_script=config["utility_dir"],
+            ignore_chr=config["ignore_chr"],
+        output:
+            fw=os.path.join(_output_dir,"{sample}.fw.bed"),
+            rv=os.path.join(_output_dir,"{sample}.rv.bed")
+        shell:
+            "python {params.dir_script}/1.1_bed2fr.py -b {input} {params.ignore_chr}"
+else:
+    rule bed2fr:
+        input:
+            os.path.join(_output_dir,"{sample}.chromapx.bed")
+        threads:
+            _threads
+        params:
+            dir_script=config["utility_dir"],
+            ignore_chr=config["ignore_chr"],
+        output:
+            fw=os.path.join(_output_dir,"{sample}.fw.bed"),
+            rv=os.path.join(_output_dir,"{sample}.rv.bed")
+        shell:
+            "python {params.dir_script}/1.1_bed2fr.py -b {input} {params.ignore_chr}"    
+
+rule bed2bdg_fw:
+    input:
+        os.path.join(_output_dir,"{sample}.fw.bed")
+    threads:
+        _threads
+    params:
+        gl=config["genomelen"]
+    output:
+        temp(os.path.join(_output_dir,"{sample}.fw.bdg"))
+    shell:
+        "bedtools genomecov -bg -i {input} -g {params.gl} > {output}"
+
+rule bed2bdg_rv:
+    input:
+        os.path.join(_output_dir,"{sample}.rv.bed")
+    threads:
+        _threads
+    params:
+        gl=config["genomelen"]
+    output:
+        temp(os.path.join(_output_dir,"{sample}.rv.bdg"))
+    shell:
+        "bedtools genomecov -bg -i {input} -g {params.gl} > {output}"
+
+rule bdg_sort_fw:
+    input:
+        fw=os.path.join(_output_dir,"{sample}.fw.bdg")
+    threads:
+        _threads
+    output:
+        temp(os.path.join(_output_dir,"{sample}.fw.sorted.bdg"))
+    shell:
+        "bedtools sort -i {input.fw} > {output}"
+
+rule bdg_sort_rv:
+    input:
+        rv=os.path.join(_output_dir,"{sample}.rv.bdg")
+    threads:
+        _threads
+    output:
+        temp(os.path.join(_output_dir,"{sample}.rv.sorted.bdg"))
+    shell:
+        "bedtools sort -i {input.rv} > {output}"
+
+if _normalize == "True":
+    rule bdg_normalize_fw:
+        input:
+            bdg=os.path.join(_output_dir,"{sample}.fw.sorted.bdg"),
+            bed=os.path.join(_output_dir,"{sample}.fw.bed")
+        threads:
+            _threads
+        params:
+            dir_script=config["utility_dir"]
+        output:
+            temp(os.path.join(_output_dir,"{sample}.fw.scaled.bdg"))
+        shell:
+            "python {params.dir_script}/1.3_bdg_normalize_v4.0.py --bdg {input.bdg} --bed {input.bed}"
+    
+    rule bdg_normalize_rv:
+        input:
+            bdg=os.path.join(_output_dir,"{sample}.rv.sorted.bdg"),
+            bed=os.path.join(_output_dir,"{sample}.rv.bed")
+        threads:
+            _threads
+        params:
+            dir_script=config["utility_dir"]
+        output:
+            temp(os.path.join(_output_dir,"{sample}.rv.scaled.bdg"))
+        shell:
+            "python {params.dir_script}/1.3_bdg_normalize_v4.0.py --bdg {input.bdg} --bed {input.bed}"
+    
+    rule bdg2bw_fw:
+        input:
+            os.path.join(_output_dir,"{sample}.fw.scaled.bdg")
+        threads:
+            _threads
+        params:
+            gl=config["genomelen"],
+            dir_script=config["utility_dir"]
+        output:
+            os.path.join(_output_dir,"{sample}.fw.scaled.bw")
+        shell:
+            "{params.dir_script}/bedGraphToBigWig {input} {params.gl} {output}"
+    
+    rule bdg2bw_rv:
+        input:
+            os.path.join(_output_dir,"{sample}.rv.scaled.bdg")
+        threads:
+            _threads
+        params:
+            gl=config["genomelen"],
+            dir_script=config["utility_dir"]
+        output:
+            os.path.join(_output_dir,"{sample}.rv.scaled.bw")
+        shell:
+            "{params.dir_script}/bedGraphToBigWig {input} {params.gl} {output}"
+    
+    rule bwAdd:
+        input:
+            fw=os.path.join(_output_dir,"{sample}.fw.scaled.bw"),
+            rv=os.path.join(_output_dir,"{sample}.rv.scaled.bw")
+        threads:
+            _threads
+        output:
+            os.path.join(_output_dir,"{sample}." + _BinSize + ".add.bdg")
+        shell:
+            """
+            bigwigCompare --binSize {_BinSize} -p {threads} --verbose -o {output} \
+            --outFileFormat bedgraph --fixedStep \
+            --bigwig1 {input.fw} \
+            --bigwig2 {input.rv} \
+            --operation add 
+            """
+else:
+    rule bdg_reverse_rv:
+        input:
+            os.path.join(_output_dir,"{sample}.rv.sorted.bdg")
+        threads:
+            _threads
+        output:
+            temp(os.path.join(_output_dir,"{sample}.rv.sorted_r.bdg"))
+        shell:
+            "awk -F '\t' -v OFS='\t' '{{$4=-$4; print}}' {input} > {output}"
+    
+    rule bdg2bw_fw:
+        input:
+            os.path.join(_output_dir,"{sample}.fw.sorted.bdg")
+        threads:
+            _threads
+        params:
+            gl=config["genomelen"],
+            dir_script=config["utility_dir"]
+        output:
+            os.path.join(_output_dir,"{sample}.fw.raw.bw")
+        shell:
+            "{params.dir_script}/bedGraphToBigWig {input} {params.gl} {output}"
+    
+    rule bdg2bw_rv:
+        input:
+            os.path.join(_output_dir,"{sample}.rv.sorted_r.bdg")
+        threads:
+            _threads
+        params:
+            gl=config["genomelen"],
+            dir_script=config["utility_dir"]
+        output:
+            os.path.join(_output_dir,"{sample}.rv.raw.bw")
+        shell:
+            "{params.dir_script}/bedGraphToBigWig {input} {params.gl} {output}"
+
+
+
+

offtracker-2.7.8/offtracker/mapping/1.1_bed2fr_v4.5.py → offtracker-2.10.0/offtracker/utility/1.1_bed2fr.py

@@ -8,19 +8,21 @@ parser.description='这算一个小彩蛋'
 # 2022.10.21. v3.0: 文件名长度 chromap -> filtered
 # 2022.10.26. v4.0: f,r 改成 fw,rv
 # 2022.01.11. v4.5: 只取 common chromosomes (chr1-chr22, chrX, chrY, chrM)
+# 2025.06.05. v5.0: 增加 ignore_chr 选项，默认只取 common chromosomes
 
 # 单文件处理脚本，配合snakemake使用
 
 parser.add_argument("-b", "--bed", type=str, metavar="dir_bed" , required=True, help="dir of bed file")
+parser.add_argument('--ignore_chr', action='store_true', help='If not set, only chr1-chr22, chrX, chrY, chrM will be analyzed.')
 
 args = parser.parse_args()
 
 bed_file = pd.read_csv( args.bed, sep='\t', header=None)
 
-common_chr = pd.Series(['chr']*22).str[:] + pd.Series(range(1,23)).astype(str).str[:]
-common_chr = pd.concat([common_chr, pd.Series(['chrX','chrY','chrM'])]).to_numpy()
-
-bed_file = bed_file[bed_file[0].isin(common_chr)]
+if not args.ignore_chr:
+    common_chr = pd.Series(['chr']*22).str[:] + pd.Series(range(1,23)).astype(str).str[:]
+    common_chr = pd.concat([common_chr, pd.Series(['chrX','chrY','chrM'])]).to_numpy()
+    bed_file = bed_file[bed_file[0].isin(common_chr)]
 
 bed_f = bed_file[bed_file[5]=='+']
 bed_r = bed_file[bed_file[5]=='-']