主要内容

ngs读数的数量

这个例子展示了如何从配对端测序读取特征,然后将它们与基因组参考联盟策划的整个人类基因组进行比对。本例使用Genome Reference Consortium Human Build 38 patch release 12 (GRCh38.p12)作为人类基因组参考。

前提条件和数据集

此示例仅适用于UNIX®和Mac平台。从Add-On Explorer下载蝴蝶结对准器支持包的生物信息学工具箱™界面。金宝app有关详细信息,请参见BioInformatics工具箱软件支持包金宝app

此示例假定您有:

建立索引

构造一个索引,用于将读取对齐到参考Bowtie2Build..文件GCF_000001405.38_GRCh38.p12_genomic.fna含有Fasta格式的人参考基因组。bowtieIdx是参考索引文件的基本名称。这“——线程8”选项指定要更快地构建索引文件的并行线程的数量。如果从同一文件夹位置运行该示例,则无需为* .fna或* .index文件指定完整的文件路径。如果要在其他位置存储文件或从其他文件夹运行示例,请指定完整路径。

Bowtiidx =“GCF_000001405.38_GRCh38.p12_genomic.index”;buildFlag = bowtie2build ('gcf_000001405.38_grch38.p12_genomic.fna'......Bowtiidx,“——线程8”);

对齐读取参考

将配对端读取为参考Bowtie2..您可以创建Bowtie2AlignOptions对象来指定不同的选项,例如要使用的并行线程数。

选择= Bowtie2alignOptions;opt.numthreads = 8;reads1 ='hg00096_1.fastq';reads2 ='hg00096_2.fastq';bowtie2 (bowtieIdx reads1 reads2,'hg00096.sam',选择);

选择性地与感兴趣的基因对齐

SAM文件可能非常大。使用BioMap仅选择正确参考的数据。对于这个例子,考虑apoe,它是与阿尔茨海默病联系的染色体19上的基因。为Apoe创建一个较小的BAM文件以提高性能。

apoeRef ='nc_000019.10'HG38中染色体19的%参考名称BM = Biomap('hg00096.sam'“SelectReference”, apoeRef);写(bm,“HG00096.bam”'格式''bam');
警告:在标题类型'PG'中发现无效标记。忽略标签“PN: bowtie2”。警告:输入SAM文件中的读序列似乎不是根据它们与参考序列对齐的开始位置来排序的。因此,在访问读操作时,性能会下降。为了获得最大的性能,在创建BioMap对象之前,对SAM文件中的读序列进行排序。

总结阅读数

使用featurecount使用GTF文件比较每个APOE变体的转录本数量。一个完整的功能表包括在GRCh38。GFF格式的p12程序集,可以通过袖口.此示例使用基于APOE成绩单的简化GTF。APOE_gene.gtf已包含在软件中。

[exportable,summary] = featurecount('apoe_gene.gtf'“HG00096.bam”......'metafeature'“transcript_id”);
处理gtf文件apoe_gene.gtf ...处理bam文件hg00096.bam ...处理参考nc_000019.10 ... 10000读取处理... 20000读取处理... 30000读取处理... 40000读取处理...50000读已被加工... 60000读已加工... 70000读取处理...... 80000读取处理...... 90000读已加工... 100000读已加工... 110000读取处理... 120000读取处理... 130000读取已加工... 140000读已加工... 150000读已加工... 160000读已加工... 190000读取处理... 190000读取处理... 190000读取处理... 200000读取处理...... 210000读取处理。.. 220000阅读加工... 230000阅读加工... 240000阅读加工... 250000阅读加工... 260000阅读加工... 270000读已加工... 280000读取处理...... 290000读取...300000读已加工... 310000读取处理... 320000读已加工... 330000读已加工... 340000读取处理... 350000阅读处理... 360000读已加工... 370000读取处理... 380000读取处理... 390000读取处理... 400000读取处理... 410000读取处理... 420000读取处理... 430000阅读加工... 440000阅读加工... 450000读取处理... 460000读已加工... 470000读取处理... 480000读取处理... 490000读取处理... 500000读已加工...510000 reads processed ... 520000 reads processed ... 530000 reads processed ... 540000 reads processed ... 550000 reads processed ... 560000 reads processed ... 570000 reads processed ... 580000 reads processed ... 590000 reads 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