Illumina高通量测序仪测序得到的原始文件是图像文件,在进行碱基识别(base calling)和Bcl2Fastq转化后,即可得到FASTQ(简称fq)格式的原始测序序列(sequenced reads),称之为raw data或raw reads。FASTQ格式的数据中含测序序列(reads)的序列信息以及其对应的测序质量信息,其中每条reads均由四行组成(Cock P.J.A. et al. 2010),示例如下: @HWI-ST1276:71:C1162ACXX:1:1101:1208:2458 1:N:0:CGATGT NAAGAACACGTTCGGTCACCTCAGCACACTTGTGAATGTCATGGGATCCAT + #55???BBBBB?BA@DEEFFCFFHHFFCFFHHHHHHHFAE0ECFFD/AEHH 第一行是序列标识(read ID)以及相关的描述信息,以“@” 开头; 第二行即为碱基序列,长度由测序策略决定; 第三行以“+”开头,后面是序列标示符、描述信息,或者什么也不加; 第四行是测序质量值(phred),与第二行一一对应,phred值以ASCII码标记,对应的 ASCII 值减去33,即为第二行对应碱基的测序质量值。 Illumina测序标识详细信息如下: HWI-ST1276 Instrument – unique identifier of the sequencer 71 run number – Run number on instrument C1162ACXX FlowCell ID – ID of flowcell 1 LaneNumber – positive integer 1101 TileNumber – positive integer 1208 X – x coordinate of the spot. Integer which can be negative 2458 Y – y coordinate of the spot. Integer which can be negative 1 ReadNumber - 1 for single reads; 1 or 2 for paired ends N whether it is filtered - NB:Y if the read is filtered out, not in the delivered fastq file, N otherwise 0 control number - 0 when none of the control bits are on, otherwise it is an even number CGATGT Illumina index sequences