CHIP分析

bowtie2使用记录

2021-02-03  本文已影响0人  shine_9457
wget http://hgdownload.cse.ucsc.edu/goldenPath/mm10/bigZips/chromFa.tar.gz    下载参考基因组
tar -zxvf chromFa.tar.gz   解压缩
cat *.fa > mm10.fa             合并文件

解压缩后文件情况:

-rw-rw-r--. 1 yangjy yangjy 133308900 Feb 10  2012 chr10.fa
-rw-rw-r--. 1 yangjy yangjy 124524201 Feb 10  2012 chr11.fa
-rw-rw-r--. 1 yangjy yangjy 122531610 Feb 10  2012 chr12.fa
-rw-rw-r--. 1 yangjy yangjy 122830079 Feb 10  2012 chr13.fa
-rw-rw-r--. 1 yangjy yangjy 127400296 Feb 10  2012 chr14.fa
-rw-rw-r--. 1 yangjy yangjy 106124566 Feb 10  2012 chr15.fa
-rw-rw-r--. 1 yangjy yangjy 100171931 Feb 10  2012 chr16.fa
-rw-rw-r--. 1 yangjy yangjy  96887024 Feb 10  2012 chr17.fa
-rw-rw-r--. 1 yangjy yangjy  92516699 Feb 10  2012 chr18.fa
-rw-rw-r--. 1 yangjy yangjy  62660205 Feb 10  2012 chr19.fa
-rw-rw-r--. 1 yangjy yangjy 199381417 Feb 10  2012 chr1.fa
-rw-rw-r--. 1 yangjy yangjy    173142 Feb 10  2012 chr1_GL456210_random.fa
-rw-rw-r--. 1 yangjy yangjy    246592 Feb 10  2012 chr1_GL456211_random.fa
-rw-rw-r--. 1 yangjy yangjy    156713 Feb 10  2012 chr1_GL456212_random.fa
-rw-rw-r--. 1 yangjy yangjy     40149 Feb 10  2012 chr1_GL456213_random.fa
-rw-rw-r--. 1 yangjy yangjy    211123 Feb 10  2012 chr1_GL456221_random.fa
-rw-rw-r--. 1 yangjy yangjy 185755495 Feb 10  2012 chr2.fa
-rw-rw-r--. 1 yangjy yangjy 163240480 Feb 10  2012 chr3.fa
-rw-rw-r--. 1 yangjy yangjy 159638285 Feb 10  2012 chr4.fa
-rw-rw-r--. 1 yangjy yangjy     68029 Feb 10  2012 chr4_GL456216_random.fa
-rw-rw-r--. 1 yangjy yangjy    232548 Feb 10  2012 chr4_GL456350_random.fa
-rw-rw-r--. 1 yangjy yangjy     15266 Feb 10  2012 chr4_JH584292_random.fa
-rw-rw-r--. 1 yangjy yangjy    212150 Feb 10  2012 chr4_JH584293_random.fa
-rw-rw-r--. 1 yangjy yangjy    195766 Feb 10  2012 chr4_JH584294_random.fa
-rw-rw-r--. 1 yangjy yangjy      2038 Feb 10  2012 chr4_JH584295_random.fa
-rw-rw-r--. 1 yangjy yangjy 154871384 Feb 10  2012 chr5.fa
-rw-rw-r--. 1 yangjy yangjy    199935 Feb 10  2012 chr5_GL456354_random.fa
-rw-rw-r--. 1 yangjy yangjy    203378 Feb 10  2012 chr5_JH584296_random.fa
-rw-rw-r--. 1 yangjy yangjy    209914 Feb 10  2012 chr5_JH584297_random.fa
-rw-rw-r--. 1 yangjy yangjy    187895 Feb 10  2012 chr5_JH584298_random.fa
-rw-rw-r--. 1 yangjy yangjy    972095 Feb 10  2012 chr5_JH584299_random.fa
-rw-rw-r--. 1 yangjy yangjy 152731283 Feb 10  2012 chr6.fa
-rw-rw-r--. 1 yangjy yangjy 148350295 Feb 10  2012 chr7.fa
-rw-rw-r--. 1 yangjy yangjy    179510 Feb 10  2012 chr7_GL456219_random.fa
-rw-rw-r--. 1 yangjy yangjy 131989244 Feb 10  2012 chr8.fa
-rw-rw-r--. 1 yangjy yangjy 127087019 Feb 10  2012 chr9.fa
-rw-rw-r--. 1 yangjy yangjy     16631 Feb 10  2012 chrM.fa
-rw-rw-r--. 1 yangjy yangjy     40874 Feb 10  2012 chrUn_GL456239.fa
-rw-rw-r--. 1 yangjy yangjy     23450 Feb 10  2012 chrUn_GL456359.fa
-rw-rw-r--. 1 yangjy yangjy     32355 Feb 10  2012 chrUn_GL456360.fa
-rw-rw-r--. 1 yangjy yangjy     48031 Feb 10  2012 chrUn_GL456366.fa
-rw-rw-r--. 1 yangjy yangjy     42915 Feb 10  2012 chrUn_GL456367.fa
-rw-rw-r--. 1 yangjy yangjy     20629 Feb 10  2012 chrUn_GL456368.fa
-rw-rw-r--. 1 yangjy yangjy     27316 Feb 10  2012 chrUn_GL456370.fa
-rw-rw-r--. 1 yangjy yangjy     29254 Feb 10  2012 chrUn_GL456372.fa
-rw-rw-r--. 1 yangjy yangjy     32251 Feb 10  2012 chrUn_GL456378.fa
-rw-rw-r--. 1 yangjy yangjy     73849 Feb 10  2012 chrUn_GL456379.fa
-rw-rw-r--. 1 yangjy yangjy     26405 Feb 10  2012 chrUn_GL456381.fa
-rw-rw-r--. 1 yangjy yangjy     23638 Feb 10  2012 chrUn_GL456382.fa
-rw-rw-r--. 1 yangjy yangjy     39449 Feb 10  2012 chrUn_GL456383.fa
-rw-rw-r--. 1 yangjy yangjy     35961 Feb 10  2012 chrUn_GL456385.fa
-rw-rw-r--. 1 yangjy yangjy     25195 Feb 10  2012 chrUn_GL456387.fa
-rw-rw-r--. 1 yangjy yangjy     29364 Feb 10  2012 chrUn_GL456389.fa
-rw-rw-r--. 1 yangjy yangjy     25178 Feb 10  2012 chrUn_GL456390.fa
-rw-rw-r--. 1 yangjy yangjy     24118 Feb 10  2012 chrUn_GL456392.fa
-rw-rw-r--. 1 yangjy yangjy     56842 Feb 10  2012 chrUn_GL456393.fa
-rw-rw-r--. 1 yangjy yangjy     24826 Feb 10  2012 chrUn_GL456394.fa
-rw-rw-r--. 1 yangjy yangjy     21681 Feb 10  2012 chrUn_GL456396.fa
-rw-rw-r--. 1 yangjy yangjy    116758 Feb 10  2012 chrUn_JH584304.fa
-rw-rw-r--. 1 yangjy yangjy 174451931 Feb 10  2012 chrX.fa
-rw-rw-r--. 1 yangjy yangjy    343694 Feb 10  2012 chrX_GL456233_random.fa
-rw-rw-r--. 1 yangjy yangjy  93579598 Feb 10  2012 chrY.fa
-rw-rw-r--. 1 yangjy yangjy    186016 Feb 10  2012 chrY_JH584300_random.fa
-rw-rw-r--. 1 yangjy yangjy    265095 Feb 10  2012 chrY_JH584301_random.fa
-rw-rw-r--. 1 yangjy yangjy    158977 Feb 10  2012 chrY_JH584302_random.fa
-rw-rw-r--. 1 yangjy yangjy    161283 Feb 10  2012 chrY_JH584303_random.fa

合并文件
cat *.fa > mm10.fa
使用bowtie2 built建立索引:
bowtie2-build mm10.fa mm10
索引文件格式如下:

-rw-r--r--. 1 yangjy yangjy  888464705 May  2  2012 mm10.1.bt2
-rw-r--r--. 1 yangjy yangjy  663195880 May  2  2012 mm10.2.bt2
-rw-r--r--. 1 yangjy yangjy       6119 May  2  2012 mm10.3.bt2
-rw-r--r--. 1 yangjy yangjy  663195875 May  2  2012 mm10.4.bt2
-rw-r--r--. 1 yangjy yangjy  888464705 May  3  2012 mm10.rev.1.bt2
-rw-r--r--. 1 yangjy yangjy  663195880 May  3  2012 mm10.rev.2.bt2

处理信息(需要一定时间)

(chipseq) [yangjy@GSCG01 align]$ bowtie2-build mm10.fa mm10
Settings:
  Output files: "mm10.*.bt2"  # 输出的文件格式:mm10.*.bt2
  Line rate: 6 (line is 64 bytes)
  Lines per side: 1 (side is 64 bytes)
  Offset rate: 4 (one in 16)
  FTable chars: 10
  Strings: unpacked
  Max bucket size: default
  Max bucket size, sqrt multiplier: default
  Max bucket size, len divisor: 4
  Difference-cover sample period: 1024
  Endianness: little
  Actual local endianness: little
  Sanity checking: disabled
  Assertions: disabled
  Random seed: 0
  Sizeofs: void*:8, int:4, long:8, size_t:8
Input files DNA, FASTA:
  mm10.fa
Building a SMALL index
Reading reference sizes
  Time reading reference sizes: 00:00:11
Calculating joined length
Writing header
Reserving space for joined string
Joining reference sequences
  Time to join reference sequences: 00:00:09
bmax according to bmaxDivN setting: 663195875
Using parameters --bmax 497396907 --dcv 1024
  Doing ahead-of-time memory usage test
  Passed!  Constructing with these parameters: --bmax 497396907 --dcv 1024
Constructing suffix-array element generator
Building DifferenceCoverSample
  Building sPrime
  Building sPrimeOrder
  V-Sorting samples
  V-Sorting samples time: 00:00:56
  Allocating rank array
  Ranking v-sort output
  Ranking v-sort output time: 00:00:14
  Invoking Larsson-Sadakane on ranks
  Invoking Larsson-Sadakane on ranks time: 00:00:25
  Sanity-checking and returning
Building samples
Reserving space for 12 sample suffixes
Generating random suffixes
QSorting 12 sample offsets, eliminating duplicates
QSorting sample offsets, eliminating duplicates time: 00:00:00
Multikey QSorting 12 samples
  (Using difference cover)
  Multikey QSorting samples time: 00:00:00
Calculating bucket sizes
Splitting and merging
  Splitting and merging time: 00:00:00
Avg bucket size: 2.65278e+09 (target: 497396906)
Converting suffix-array elements to index image
Allocating ftab, absorbFtab
Entering Ebwt loop
Getting block 1 of 1
  No samples; assembling all-inclusive block
  Sorting block of length 2652783500 for bucket 1
  (Using difference cover)

运行bowtie2 获取 SAM 文件
bowtie2 -p 6 -3 5 --local -x mm10 -1 example_1.fastq -2 example_2.fastq -S example.sam

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