rna-seq分析—py+linux

zky___

已于 2024-04-15 13:46:23 修改

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文章标签： linux python pandas

于 2024-03-21 09:22:23 首次发布

本文链接：https://blog.csdn.net/m0_69569041/article/details/136595360

版权

太久不用了啥都忘了一步步走..

1.重新下载ubuntu就花了好久....

2.下好了以后关联python3命令到python3.9.6(习惯）



sudo ln -s /usr/bin/python3 /usr/bin/python


zky@ZKY:/mnt/g/linux_zky/Python-3.9.6$ python3
Python 3.9.6 (default, Mar 10 2024, 08:00:53)
[GCC 9.4.0] on linux
Type "help", "copyright", "credits" or "license" for more information.
>>> quit()

3.下载

apt install hisat-2
conda install -c bioconda stringtie

4.建库

hisat2-build --bmax 389309110 --dcv 1024 -p 8 genome.fa genome

5.文件位置

/mnt/e/转录组全部原始数据/DZOE2024011942-b1

#双端文件比对mapping到参考基因组

hisat2 -t -p 20 -x /mnt/g/linux_zky/database/genome \ -1 H_S_L_l_1_R1.fq -2 H_S_L_l_1_R2.fq -S /mnt/g/linux_zky/H_S_L_l_1.sam

排序

 samtools view -@ 16 -S H_S_L_l_1.sam -b > H_S_L_l_1.bam

samtools sort H_S_L_l_1.bam -o H_S_L_l_1_sort.bam -@ 16

单个样本组装

stringtie -p 16 H_S_L_l_1_sort.bam -G genes.gtf -o H_S_L_l_1.gtf

这里通过脚本进行批量单个样本的组装

import os
import subprocess

# 项目位置：/mnt/g/linux_zky/

# 定义目录路径
directory = "/mnt/e/转录组全部原始数据/DZOE2024011942-b1"

unique_list = set()
for filename in os.listdir(directory):
	# 构建完整的文件路径
	print(filename.split(".")[0])
	
	unique_list.add(filename.split(".")[0])

print(unique_list.__len__())

for i in unique_list:
	print("run cmd:", i)
	#这里重新加了-e 不加就会获得所有转录本 包括未知的
	cmd4 = f'stringtie -e -p 64 /mnt/g/linux_zky/bam_gtfs/bams/{i}_sort.bam -G /mnt/g/linux_zky/genes.gtf -o /mnt/g/linux_zky/test/sort_gtf/{i}_sort.gtf'
	subprocess.run(cmd4, shell=True)

获得strimerge.gtf

1.emergeLIst

import os

# 项目位置：/mnt/g/bam_gtfs/gtfs

# 定义目录路径
directory = "/mnt/g/linux_zky/test/sort_gtf/"
allGtfs = open("mergelist.txt", "w")

unique_list = set()
for filename in os.listdir(directory):
	# 构建完整的文件路径
	allGtfs.write(directory + filename + "\n")

#获得strimerge.gtf
"""
then=>
stringtie --merge -p 16 -G genes.gtf -o strimerge.gtf mergelist.txt
"""

2.组装

stringtie --merge -p 16 -G genes.gtf -o strimerge.gtf mergelist.txt

perpDE脚本下载

Ballgown获得所有转录本及其丰度

import os
import subprocess
# "/mnt/g/linux_zky/test/ballGown/b_gtfs"

# 定义目录路径
directory_o_gtfs = "/mnt/g/linux_zky/test/ballGown/b_gtfs/"
directory_o_tabs = "/mnt/g/linux_zky/test/ballGown/b_tabs/"
directory_in = "/mnt/g/linux_zky/bam_gtfs/bams/"

for filename in os.listdir(directory_in):
	# 构建完整的文件路径
	
	sortBam = directory_in + filename
	oriName = filename.split("_sort")[0]
	ballgown_gtf = directory_o_gtfs + oriName + ".gtf"
	gene_abund = directory_o_tabs + oriName + ".tab"
	
	print(sortBam, gene_abund, ballgown_gtf)
	
	cmd = f'stringtie -e -B -p 64 -G strimerge.gtf -o {ballgown_gtf} {sortBam} -A {gene_abund}'
	subprocess.run(cmd, shell=True)

命令行：

python prepDE.py3 -i /mnt/g/bam_gtfs/rnaSeqAnalyse/sample_list.txt -g /mnt/g/bam_gtfs/rnaSeqAnalyse/gene_count.csv -t /mnt/g/bam_gtfs/rnaSeqAnalyse/transcript.csv

sample_list

# 代替字典
replace_dict = {
	"H": "12h",
	"W": "6d",
	"M": "13d",
	"S": "9782",
	"T": "9614",
	"N": "np",
	"L": "lp",
	
}

import os

# 定义目录路径
directory = "/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/"
all_sample = open("sample_list.txt", "w")

for filename in os.listdir(directory):
	# 构建完整的文件路径
	
	if filename.split(".")[1] == "gtf":
		file = directory + filename
		sample = filename.split(".")[0]
		for key in replace_dict:
			sample = sample.replace(key, replace_dict[key])
		
		print(sample)
		print(file)
		all_sample.write(sample + "\t" + directory + filename + "\n")

"""
(zky) zky@ZKY:/mnt/g/linux_zky/bam_gtfs/stringtie-master$
python prepDE.py3 -i /mnt/g/linux_zky/mapping后分析/sample_list.txt -g /mnt/g/linux_zky/mapping后分析/preDEdata/gene_count.csv -t /mnt/g/linux_zky/mapping后分析/preDEdata/transcript.csv"""

欧易云工具

sample_list

12h_9782_lp_l_1	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_l_1.gtf
12h_9782_lp_l_2	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_l_2.gtf
12h_9782_lp_l_3	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_l_3.gtf
12h_9782_lp_r_1	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_r_1.gtf
12h_9782_lp_r_2	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_r_2.gtf
12h_9782_lp_r_3	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_L_r_3.gtf
12h_9782_np_l_1	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_N_l_1.gtf
12h_9782_np_l_2	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_N_l_2.gtf
12h_9782_np_l_3	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_N_l_3.gtf
12h_9782_np_r_1	/mnt/g/linux_zky/mapping后分析/ballGown/b_gtfs/H_S_N_r_1.gtf
............

prepDE:

(zky) zky@ZKY:/mnt/g/linux_zky/bam_gtfs/stringtie-master$ 
python prepDE.py3 
-i /mnt/g/linux_zky/mapping后分析/sample_list.txt 
-g /mnt/g/linux_zky/mapping后分析/preDEdata/gene_count.csv 
-t /mnt/g/linux_zky/mapping后分析/preDEdata/transcript.csv

重新调整

import pandas as pd

g_c = pd.read_csv("G:\linux_zky\mapping后分析\preDEdata\gene_count.csv")

test = g_c[~g_c['gene_id'].str.contains("\|")]
print(test)
g_c['gene_id'] = g_c['gene_id'].str.split("\|").str[1]

g_c.to_csv("G:\linux_zky\mapping后分析\preDEdata\gene_count_2.csv", index=False)  # 写入新的 CSV 文件，不包含索引列

重排序所有tab

import os

import pandas as pd
#排序用于整理出表达量矩阵
dictionary = r"G:\linux_zky\mapping后分析\ballGown\b_tabs"

for i in os.listdir(dictionary):
	with open(os.path.join(dictionary, i), "r") as f:
		reader = pd.read_csv(f, sep='\t')
		print(i)
		i_index = reader.sort_values(by="Gene Name")
		i_index.iloc[:,1:].to_csv(fr"G:\linux_zky\mapping后分析\ballGown\sorted\{i}_sort.csv", index=False)  # 写入新的 CSV 文件，不包含索引列

用重排的tabs自己实现表达量矩阵

import os

import pandas as pd

replace_dict = {
	"H": "12h",
	"W": "6d",
	"M": "13d",
	"S": "9782",
	"T": "9614",
	"N": "np",
	"L": "lp",
	
}

dictionary = r"G:\linux_zky\mapping后分析\ballGown\sorted"
all_FPKM_values = []  # 用于存储所有的 FPKM 值
with open(r"G:\linux_zky\mapping后分析\ballGown\sorted\H_S_L_l_1.tab_sort.csv", "r") as f_head:
	reader = pd.read_csv(f_head, sep=',')
	FPKM_head = reader.iloc[:, :1]  # 获取 FPKM 列
	all_FPKM_values.append(FPKM_head)
df = pd.concat(all_FPKM_values, axis=1)  # 将多个 FPKM 列合并为一个 DataFrame


for i in os.listdir(dictionary):
	if i.endswith(".csv"):
		with open(os.path.join(dictionary, i), "r") as f:
			reader = pd.read_csv(f, sep=',')
			
			FPKM_column = reader.iloc[:, 7]  # 获取 FPKM 列
			
			i = i.split(".")[0]
			for key in replace_dict:
				i = i.replace(key, replace_dict[key])

			df[i] = FPKM_column

df.to_csv("fpkm_matrix.csv", index=False)  # 写入新的 CSV 文件，不包含索引列

g_diff_DEG_group.py


from openpyxl import Workbook

wb = Workbook()
ws = wb.active
ws.append(["case", "case_name", "control", "control_name", "replicate", "paired", "method"])

group_ = [
    "12h_9782_l",
    "12h_9782_r",
    "12h_9614_l",
    "12h_9614_r",
    "6d_9782_l",
    "6d_9782_r",
    "6d_9614_l",
    "6d_9614_r",
    "12d_9782_l",
    "12d_9782_r",
    "12d_9614_l",
    "12d_9614_r",
]
days = ["12h", "6d", "13d"]
materials = ["_9782", "_9614"]
c_c = ["_lp", "_np"]
parts = ["_l", "_r"]
repetition = ["_1", "_2", "_3"]

for day in days:
    for material in materials:
        for part in parts:
            case = day + material + c_c[0] + part + repetition[0] + "," + day + material + c_c[0] + part + repetition[1] + "," + day + material + c_c[0] + part + repetition[2]
            caseName = day + material + c_c[0] + part
            control = day + material + c_c[1] + part + repetition[0] + "," + day + material + c_c[1] + part + repetition[1] + "," + day + material + c_c[1] + part + repetition[2]
            controlName = day + material + c_c[1] + part
            replicate = "yes"
            paired = "yes"
            method = "DESeq2"
            ws.append([case, caseName, control, controlName, replicate, paired, method])

wb.save("diff_deg_group.xlsx")

用欧易云做吧做了degs分析

欧易云的分析工具

——————
——

wgcna分析

由于进行WGCNA分析时候组别太多并且全部的表达量矩阵会导致出来的聚类模块非常错乱

并且在三个样本里面均质性不好

所以就用三个生物学重复在DEGS里面验证过P值Q值的基因直接提出来分别拿到根和叶中的表达量矩阵

重做表达量矩阵....

"""

将把表达量矩阵重做成平均值来分析
将每组三列数据的平均值作为新的一列，且新列的名称保留原始列名的前缀部分。
"""

import pandas as pd

f_ubundant = pd.read_csv("newFpkmMatrix.csv", index_col=0)

# 提取每组三列数据的前缀部分，例如将 '12h_9614_lp_l_1' 转换为 '12h_9614_lp_l'
column_prefixes = set(col.rsplit('_', 1)[0] for col in f_ubundant.columns[:])
print(len(column_prefixes))
# 计算每组三列数据的平均值
averaged_data = pd.DataFrame()
for prefix in column_prefixes:
	cols = [col for col in f_ubundant.columns if col.startswith(prefix)]
	print(cols)
	averaged_data[prefix] = round(f_ubundant[cols].mean(axis=1), 1)
	print(prefix)
# 保存结果
averaged_data.to_csv('averaged_gene_FPKM.csv')

a获取了表达量均值的矩阵后面就是把DEGS差异文件里面的每一个-diff-p-val-0.05-FC-2.gene.xls文件的相应组别的基因号提取出来然后去除表达量太低的 (<1.5)

返回对照到矩阵就好了

import os

import pandas as pd

id_L = set()
id_R = set()

for filename in os.listdir("../DEGS"):
	if filename.endswith("0.05-FC-2.gene.xls"):
		if "_l-" in filename:
			f_l = pd.read_csv("../DEGS/" + filename, sep="\t")
			id_L.update(f_l['gene_id'])  # 使用 update() 将 'gene_id' 列的值添加到集合中
		elif "_r-" in filename:
			f_l = pd.read_csv("../DEGS/" + filename, sep="\t")
			id_R.update(f_l['gene_id'])  # 使用 update() 将 'gene_id' 列的值添加到集合中
		else:
			pass

# 读取表达量矩阵
matrix = pd.read_csv("averaged_gene_FPKM.csv")

# 获取包含 "_l" 的列名
l_columns = [col for col in matrix.columns if "p_l" in col]

# 获取包含 "_r" 的列名
r_columns = [col for col in matrix.columns if "_r" in col]

# 根据 id_L 筛选行,并选择 "_l" 列创建 f_lMatrix
f_lMatrix = matrix[matrix["gene_id"].isin(id_L)][["gene_id"] + l_columns]

# 去除所有基因表达量都低于1.5的基因
f_lMatrix = f_lMatrix[(f_lMatrix[l_columns] > 1.5).any(axis=1)]
f_lMatrix.drop_duplicates(subset="gene_id",inplace=True)
f_lMatrix.to_csv("f_lMatrix.csv", index=False)

# 根据 id_R 筛选行,并选择 "_r" 列创建 f_rMatrix
f_rMatrix = matrix[matrix["gene_id"].isin(id_R)][["gene_id"] + r_columns]

# 去除所有基因表达量都低于1.5的基因
f_rMatrix = f_rMatrix[(f_rMatrix[r_columns] > 1.5).any(axis=1)]
f_rMatrix.drop_duplicates(subset="gene_id",inplace=True)
f_rMatrix.to_csv("f_rMatrix.csv", index=False)

gene_id,12h_9782_lp_l,13d_9614_lp_l,6d_9782_np_l,6d_9782_lp_r,13d_9614_np_r,6d_9782_np_r,12h_9614_np_r,13d_9782_lp_r,13d_9614_lp_r,6d_9614_lp_r,6d_9614_lp_l,13d_9782_lp_l,12h_9782_lp_r,12h_9782_np_l,12h_9614_np_l,12h_9782_np_r,13d_9782_np_l,6d_9782_lp_l,6d_9614_np_r,12h_9614_lp_l,6d_9614_np_l,13d_9782_np_r,12h_9614_lp_r,13d_9614_np_l
Zm00001d014914,29.3,14.9,26.4,0.8,0.8,0.4,0.7,0.3,1.1,12.2,16.8,21.3,2.1,14.6,22.5,0.1,4.2,22.9,1.9,20.9,3.9,0.4,0.9,6.9
Zm00001d023904,33.4,4.6,36.5,31.9,5.2,35.9,8.0,35.4,4.9,6.3,5.1,34.6,38.0,39.2,5.0,52.0,28.8,40.6,5.9,4.3,5.2,35.0,7.0,5.5
Zm00001d007951,52.1,73.8,82.9,83.1,98.8,84.7,108.4,98.0,111.4,84.7,65.0,78.8,76.2,44.7,36.4,110.2,68.2,83.1,87.8,32.4,66.1,79.1,102.8,71.6
Zm00001d006372,0.1,0.1,0.2,0.9,0.1,0.9,0.7,0.1,0.0,0.5,0.1,0.1,0.3,0.0,0.0,0.2,0.3,0.3,1.5,0.0,0.0,0.3,0.5,0.0
Zm00001d034644,279.8,314.2,381.5,595.8,448.5,581.4,402.1,492.6,485.1,478.5,272.4,345.7,499.2,254.9,283.1,450.9,337.5,349.5,460.0,266.8,280.9,604.6,451.5,310.1

传入相关性状....

wgcna分析

————————

分析补充1：玉米的B73数据库+基因注释——匹配整理

要求对照

的基因id，直接在https://www.ncbi.nlm.nih.gov/gene搜了

(b73) AND "Zea mays"[porgn:__txid4577]

把列表信息全下下来了

b73_geneData.xls

Aliases	description	other_designations	map_location	chromosome	genomic_nucleotide_accession.version	start_position_on_the_genomic_accession	end_position_on_the_genomic_accession	orientation	exon_count	OMIM
ZEAMMB73_Zm00001d001812, GRMZM2G457621, RAF1	Rubisco accumulation factor 1, chloroplastic	rubisco accumulation factor 1, chloroplastic\|Rubisco Assembly Factor 1		2	NC_050097.1	1617667	1619220	plus	1
ZEAMMB73_Zm00001d044122, DFR, FNR, GRMZM2G026930, pco110172(456)	Dihydroflavonol 4-reductase	dihydroflavonol 4-reductase\|A1 (Teosinte)\|Dihydrokaempferol 4-reductase\|NADPH-dependent reductase\|NADPH-dependent reductase-like protein\|dihydroflavonol4-reductase\|flavanone 4-reductase		3	NC_050098.1	221779797	221781573	minus	4
ZEAMMB73_Zm00001d050133, GRMZM2G365423, GRMZM2G389303, GRMZM2G437977, umc1175	Bifunctional aspartokinase/homoserine dehydrogenase 1, chloroplastic	bifunctional aspartokinase/homoserine dehydrogenase 1, chloroplastic\|AK-HD 1\|AK-HSDH 1\|Bifunctional aspartokinase/homoserine dehydrogenase 2 chloroplastic\|Homoserine dehydrogenase\|aspartate kinase-homoserine dehydrogenase		4	NC_050099.1	68464991	68510882	minus	19
ZEAMMB73_Zm00001d050339, ALDH10A8, AMADH1, GRMZM2G013214, GRMZM2G146754, ZmAMADH1a, gpm154	Aminoaldehyde dehydrogenase 1a	aminoaldehyde dehydrogenase 1a\|4-trimethylammoniobutyraldehyde dehydrogenase AMADH1a\|Aminobutyraldehyde dehydrogenase AMADH1a\|Betaine aldehyde dehydrogenase 2 mitochondrial\|Betaine aldehyde dehydrogenase AMADH1a\|Gamma-guanidinobutyraldehyde dehydrogenase AMADH1a\|aminoaldehyde dehydrogenase 1		4	NC_050099.1	82466139	82471927	plus	15

后面用正则慢慢配到Symbol这一列

"""
输入任意表头的单列基因编号文件 用于匹配相应基因注释
"""

import pandas as pd

# 读取基因文件
f_geneData = r'b73T1.csv'
f_geneData = pd.read_csv(f_geneData)

csv_ = pd.read_csv("input.txt")
print(csv_)

result_description = []
result_aliases = []

for i in csv_.iloc[:, 0]:
	
	target = 'description'
	f_g_t_aliases = f_geneData[f_geneData['Aliases'].str.contains(i)]
	if len(f_g_t_aliases) != 0:
		description = f_g_t_aliases[target].values[0]
		aliases = f_g_t_aliases['other_designations'].values[0]
		result_description.append(description)
		result_aliases.append(aliases)
	else:
		f_g_t_symbol = f_geneData[f_geneData['Symbol'].str.contains(i)]
		if len(f_g_t_symbol) != 0:
			description = f_g_t_symbol[target].values[0]
			aliases = f_g_t_symbol['other_designations'].values[0]
			result_description.append(description)
			result_aliases.append(aliases)
		else:
			print("没有匹配到:", i)
			result_description.append(None)
			result_aliases.append(None)

csv_['description'] = result_description
# csv_['Aliases'] = result_aliases
print(csv_)
csv_.to_csv("结果.csv", index=False)

分析补充2 ：自己计算所有基因的log2FoldChange

"""
对这样的一个csv文件写一个脚本 首先
取表头的前半部分和最后一部分比如 12h_9782_lp_l 取到的应该是 12h_9782_l 可以考虑用split("_")进行拼接 新建一个dataframe，
之后按照set中的内容分别检索 比如12h_9782_l是一个set中的元素 取12h_9782_np_l 和 12h_9782_lp_l那么往这个新的df中加入一列，
命名为12h_9782_l-LPvsNP 其中的内容是12h_9782_lp_l/12h_9782_np_l 的值 最后生成一个含有12个纵列的比值df 存为csv ```

在修改后的代码中，我们定义了一个伪计数pseudocount，其值为0.001。在计算log2FoldChange之前，我们将处理组和对照组的表达量值分别加上伪计数。

result_df[f"{col}-LPvsNP"] = np.log2((df[lp_col] + pseudocount) / (df[np_col] + pseudocount))

通过添加伪计数，我们可以避免除以0或计算log2(0)的问题。这种方法在处理RNA-seq或其他高通量数据时常用，可以有效地处理表达量为0的情况。

请注意，伪计数的选择可能会影响结果的解释。选择一个合适的伪计数值取决于数据的特点和研究目的。常用的伪计数值包括0.001、1e-5或1。
"""
import pandas as pd

# 读取CSV文件
df = pd.read_csv('averaged_gene_FPKM.csv')

# 提取列名的第1、2、4个元素
columns = df.columns[1:]
column_set = set(["_".join(col.split("_")[:2] + [col.split("_")[3]]) for col in columns])

# 创建一个新的DataFrame用于存储比值
result_df = pd.DataFrame()

# 将gene_id添加到结果DataFrame中
result_df['gene_id'] = df['gene_id']

# 遍历集合中的每个元素
for col in column_set:
	lp_col = [c for c in columns if col.split("_")[0] in c and col.split("_")[1] in c and col.split("_")[2] in c and "lp" in c]
	np_col =[c for c in columns if col.split("_")[0] in c and col.split("_")[1] in c and col.split("_")[2] in c and "np" in c]
	
	if lp_col and np_col:
		lp_col = lp_col[0]
		np_col = np_col[0]
		
		# 计算比值并添加到结果DataFrame中
		result_df[f"{col}-LPvsNP"] = df[lp_col] / df[np_col]

result_df = result_df.round(3)
# 保存结果为新的CSV文件
result_df.to_csv('output.csv', index=False)

这种方法可以获得差异倍数文件：
用于表达量作图自定义基因的表达差异分析

gene_id,6d_9782_l-LPvsNP,6d_9782_r-LPvsNP,6d_9614_r-LPvsNP,6d_9614_l-LPvsNP,13d_9614_l-LPvsNP,12h_9782_l-LPvsNP,13d_9782_l-LPvsNP,12h_9614_l-LPvsNP,12h_9614_r-LPvsNP,12h_9782_r-LPvsNP,13d_9782_r-LPvsNP,13d_9614_r-LPvsNP
Zm00001d014914,-5.0426,0.9982,2.6822,1.6451,1.1105,1.0049,-3.8029,-0.1064,0.3621,4.3786,-0.4138,0.4589
Zm00001d023904,-0.1943,-0.1704,0.0946,0.2768,-0.2577,-0.231,0.2977,-0.2175,-0.1926,-0.4525,0.0164,-0.0857
Zm00001d007951,0.0035,-0.0275,-0.0519,0.3577,0.0437,0.221,0.523,-0.1679,-0.0765,-0.5323,0.3091,0.1732
Zm00001d006372,2.1643,0.0,-1.583,8.9687,6.6582,6.6582,-1.5754,0.0,-0.4846,0.5826,-1.5754,-6.6582
Zm00001d034644,0.6431,0.0353,0.0569,0.7685,0.0189,0.1345,0.5455,-0.0856,0.1672,0.1468,-0.2956,0.1132
Zm00001d016572,-0.3539,-0.0895,0.089,0.4088,-0.037,0.7711,0.3268,-0.5075,0.1355,0.1817,0.2278,-0.2194
Zm00001d017857,0.2529,-0.085,0.03,1.1262,-0.1072,0.0842,0.8826,-0.5577,0.5288,0.1322,0.1466,0.9272
Zm00001d051685,-0.1389,-0.1106,-0.0252,1.0196,-0.0179,0.0366,0.634,-0.5733,0.4165,-0.0442,0.1448,0.6312
Zm00001d040880,0.5826,-0.735,-0.5826,0.0,0.0,0.4138,0.5826,0.0,0.0,-1.3176,0.0,1.3176

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rna-seq分析—py+linux

太久不用了啥都忘了一步步走..1.重新下载ubuntu就花了好久....2.下好了以后关联python3命令到python3.9.6(习惯）3.下载4.建库5.文件位置/mnt/e/转录组全部原始数据/DZOE2024011942-b1#双端文件比对mapping到参考基因组排序。
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