Bowtie2 aligner not generating .sam file.
2
0
Entering edit mode
6.4 years ago
Ramana • 0

Hi, I used bowtie2 V2.3.3.1 1st Step- bowtie2-build -f /media//CANCERGENOMICS/NewFolder/hg38.fa hg38 Generated a list of files- hg38.1.bt2, hg38.2.bt2,hg38.3.bt2,hg38.4.bt2,hg38.fa, hg38.rev.1.bt2, hg38.rev.2.bt2

2nd Step- bowtie2 -x hg38 -1 /media/CANCERGENOMICS/NewFolder/bowtie2/SRR098401testsickle_1.fastq -2 /media//CANCERGENOMICS/NewFolder/bowtie2/SRR098401testsickle_2.fastq -s /media/CANCERGENOMICS/NewFolder/bowtie2/SRR098401_pebwt.sam

After executing cmd it running for long time.

Result - I'm not able to generate the file "SRR098401_pebwt.sam" (?) The prompt shows( last few lines) like this..... Can any one explain me why it is happening ?

SRR098401.114048463 147 chr9 123652773 8 28M = 123652716 -85 AAAAAAAACGCAAAACAAACAGCAGCAA @@@AACCC6,(<;7FFGGHHHCHHBHHH AS:i:-13 XN:i:0 XM:i:4 XO:i:0 XG:i:0 NM:i:4 MD:Z:8T0T0G2G14 YS:i:-14 YT:Z:CP 92522058 reads; of these: 92522058 (100.00%) were paired; of these: 2035529 (2.20%) aligned concordantly 0 times 73726342 (79.69%) aligned concordantly exactly 1 time 16760187 (18.11%) aligned concordantly >1 times ---- 2035529 pairs aligned concordantly 0 times; of these: 949789 (46.66%) aligned discordantly 1 time ---- 1085740 pairs aligned 0 times concordantly or discordantly; of these: 2171480 mates make up the pairs; of these: 992625 (45.71%) aligned 0 times 387998 (17.87%) aligned exactly 1 time 790857 (36.42%) aligned >1 times 99.46% overall alignment rate SRR098401.114048513 99 chr10 132415511 3 45M1I4M = 132415574 101 ATCCAGACATCCACTGTCACACACACGCACTCTCACACACATTCACCACC HGHFHFHHEFHCHHHHEHEHHFEBHFHHFHHHEHHDHHGHCCEFEAC7BD AS:i:-13 XN:i:0XM:i:1 XO:i:1 XG:i:1 NM:i:2 MD:Z:48A0 YS:i:-21 YT:Z:CP SRR098401.114048513 147 chr10 132415574 3 38M = 132415511 -101 CACAACCACCCACACACACCCACCCACAGACACCCACA @<c@2?b>D.DAD@ECE;BBD>D;5<=:-.8>/C;CB AS:i:-21 XS:i:-15 XN:i:0 XM:i:7XO:i:0 XG:i:0 NM:i:7 MD:Z:4T4T13T4T0T0T2T4 YS:i:-13 YT:Z:CP SRR098401.114048581 83 chr10 1816393 24 45M = 1816267 -171 TTTGACTGATATTTGTATAGCCACCGCAGCTCTCTTTTGGTTACT EEDD?CAA>DEBFEBEBDBFFF?AF?GFAG@GGGGGGGGGGGFFG AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:45 YS:i:-13 YT:Z:CP SRR098401.114048581 163 chr10 1816267 24 23M = 1816393 171 CGTTTGGTTCTTTTTTTTTGGTG @HHHHGDD9@GGEA?FFF7F6:; AS:i:-13 XN:i:0 XM:i:3 XO:i:0 XG:i:0 NM:i:3 MD:Z:0A7A9A4 YS:i:0 YT:Z:CP SRR098401.114048604 97 chr6 98176005 14 21M1D24M chr17 77085279 0 AATTGAAGGAAATTGAAATATAAAAAAAAAAATTCAAAAGATCAA HHHHHHHHHHHHHHHHHHHEHHHGHHHHHHE6G?:DGD?A2::B= AS:i:-8 XN:i:0 XM:i:0 XO:i:1XG:i:1 NM:i:1 MD:Z:21^A24 YT:Z:UP SRR098401.114048604 145 chr17 77085279 42 25M chr6 98176005 0 AAAAAAAATAAAACCCAAAAAAAGA @@@@-<<<&?56@@EEEE7CCC@C AS:i:-2 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:8A16 YT:Z:UP SRR098401.114048637 99 chr15 33685784 7 50M = 33685888 124 AAACTCACTCAAAACCGCACAACTACATGGAAACCGAACAATCTGCTCCT EDBGGGGGFEGGGGGGGGGGGGGEGGGFGFGGBGGGAEDCGGGGG7DCDC AS:i:0 XS:i:-5 XN:i:0XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:50 YS:i:-11 YT:Z:CP SRR098401.114048637 147 chr15 33685888 7 20M = 33685784 -124 AACCAAAAAAAACAAAAACA CBA@@<%.<4;<;6<>-@=> AS:i:-11 XS:i:0 XN:i:0 XM:i:4 XO:i:0 XG:i:0 NM:i:4 MD:Z:6T0G1G6G3 YS:i:0YT:Z:CP SRR098401.114048738 89 chr3 175315939 42 45M = 175315939 0 TTCATATATGACCCTATATTTAAAAATGCAATTAATGTGTCTTTG EEEBEGEGGGFBEEGGDGGBGFFGGGGGFGGGEGGGGFGGGGGFG AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0NM:i:0 MD:Z:45 YT:Z:UP SRR098401.114048738 165 chr3 175315939 0 * = 175315939 0 TGTTTGTGTTTTTTTTTTGTTGTGG C8@AA1<>@<;)<;@1@1@<<9?9 YT:Z:UP SRR098401.114048748 73 chr7 20086531 15 54M = 20086531 0 CCACACCCAGCTAATTTTTGTATTATTAGTAGAGACAAGGTTTTACCCTGTTGG HHHHHHHGGHHGGGHHGHHHBEGBEGGFGFHGDHDHHDHHEHHHF@@B@@>B@@ AS:i:-5 XN:i:0XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:47A6 YT:Z:UP SRR098401.114048748 133 chr7 20086531 0 * = 20086531 0 GGGTGGTTTTTTTGGGGTTGGGTGT HHHHH@2C24-<<<1?8894CB9B? YT:Z:UP SRR098401.114048920 89 chr5 70946170 1 45M = 70946170 0 GTATATGGTAAGTAATCACTCAGCATCTTTTCCTGACAATTTTTT ?EGGFDHHHHHHHHHHHHHHHHGHHHBHHHHHHHHHHEHHHHHHH AS:i:-5 XN:i:0 XM:i:1 XO:i:0 XG:i:0NM:i:1 MD:Z:0T44 YT:Z:UP SRR098401.114048920 165 chr5 70946170 0 * = 70946170 0 TCGGGGGTTTGGTTTTTTGTTTGGTGG @,68@02@@@21A@ADDBBBBB%8D=> YT:Z:UP SRR098401.114048974 97 chr20 43139895 42 45M chr7 39680385 0 AAAGCCTCCAAGGGACTTACCACAAAACAGAAGCAATATTTTAAC FEEDEBGGGAAFFB@EFEEEFGFF?FDGEE>@;DC;:DDAD=?DB AS:i:-5 XN:i:0 XM:i:1 XO:i:0 XG:i:0NM:i:1 MD:Z:44A0 YT:Z:UP SRR098401.114048974 145 chr7 39680385 1 20M chr20 43139895 0 ACAAAAACAAAAAAAACACA ?79??<@<<4471B>5-5@@ AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:20 YT:Z:UP SRR098401.114048986 89 chr10 84627148 42 38M = 84627148 0 CCTGCCATTATGTTACATGATGTTCTTTGCCTGTTACA EEB?HHHFFGFFCBEEFHHAHFDGHGFHEGFGFGFFGG AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0MD:Z:38 YT:Z:UP SRR098401.114048986 165 chr10 84627148 0 * = 84627148 0 GGGTTTTTTTGGTTGGTTTGTGTGTGGTGTTTTTTG ??=643?>;<.41;?=94?<>>;.@BA:1AGDBB?: YT:Z:UP SRR098401.114049008 99 chrX 25575707 40 44M = 25575835 148 TGGGAATAAGAGGCTAATAGGAGAGGACAAGTAGTGCCAAAGTA HHHHFHHHHHHGHHHHDHHHEHGEHHFHHHGCGFBFFFED<c6b as:i:0="" xn:i:0="" xm:i:0="" xo:i:0="" xg:i:0nm:i:0="" md:z:44="" ys:i:-11="" yt:z:cp="" srr098401.114049008="" 147="" chrx="" 25575835="" 40="" 20m="25575707" -148="" aaaaaaacaaaaacaaaaaa="" @@@@@@'-@@ee7eegg(gd="" as:i:-11="" xs:i:0="" xn:i:0="" xm:i:4="" xo:i:0="" xg:i:0="" nm:i:4="" md:z:6t0g4g4t2="" ys:i:0yt:z:cp="" srr098401.114049024="" 83="" chr2="" 51191331="" 8="" 45m="51191234" -142="" gttgttaacttgagatatttctagcttttcagtgtgggcaattag="" bfbgfhhhhfhbhhhhhhghhfhhhhhhhhhhhhhhhhhhhhghh="" as:i:-5="" xs:i:-20="" xn:i:0="" xm:i:1xo:i:0="" xg:i:0="" nm:i:1="" md:z:0t44="" ys:i:-33="" yt:z:cp="" srr098401.114049024="" 163="" chr2="" 51191234="" 8="" 69m="51191331" 142="" tcttgtgttttggtggttttggggtttgtttgtttttggttttgttttggtttggtttgttgttggttg="" c,ccc@ccc2@;4="">/A4CCAEGGECHHHFH==2?2?=?@@A'@9=;<7'>?CC'C?CAC.@%BBA>>@> AS:i:-33 XN:i:0 XM:i:12 XO:i:0 XG:i:0 NM:i:12 MD:Z:9C2A1A1C15C1C6C1C4A4A5C1C7 YS:i:-5 YT:Z:CP SRR098401.114049028 147 chr9 63254389 1 32M = 63254344 -77 CAAAACAAAACACACACCCCCACACAAACACA 7;@?@%@EGDGDFCFEEGFFGFHHGHHHHHHH AS:i:-2 XS:i:-2 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1MD:Z:5T26 YS:i:0 YT:Z:CP SRR098401.114049061 89 chr2 106552997 42 45M = 106552997 0 GGCACTGTAATATTTTGGGAATCTTGCTTTTCTGCACTTTTCCCT BFGGCGHHFHEGGGGGECGHHHHGGHHDHHEGGEGDFFHGHHHHG AS:i:-5 XN:i:0 XM:i:1 XO:i:0 XG:i:0NM:i:1 MD:Z:17T27 YT:Z:UP SRR098401.114049061 165 chr2 106552997 0 * = 106552997 0 TTGTGGTTGGTTATGTTTTGTGT <,::?74=6<<,?;@@B<4??A YT:Z:UP SRR098401.114049095 113 chr12 104805213 42 50M chr17 30910349 0 AACAGAGCTGTCCCGAGCAGATCACCCAATGCTGTTAGGTAGGGGATGGA 89;68EFFEEFCBF?FFEFFB<ecefe@fefff?fffffffff@faefff as:i:0="" xn:i:0="" xm:i:0xo:i:0="" xg:i:0="" nm:i:0="" md:z:50="" yt:z:up="" srr098401.114049095="" 177="" chr17="" 30910349="" 1="" 21m="" chr12="" 104805213="" 0="" accaaaaaaaaaaaaacacca="" @c?@@24&lt;:'7;@@5="">7>81: AS:i:-2 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:9C11 YT:Z:UP SRR098401.114049150 65 chr11 101643970 0 42M chr8 73321970 0 AGGCATGGGCAAGGACTTCATGACTACCGCACGAAAAGCAAT FEDGGFGBDGGGFFCFFFFDGBFDGB;<<:<<<<<eb8ed@c as:i:-16="" xn:i:0="" xm:i:4="" xo:i:0xg:i:0="" nm:i:4="" md:z:26a0a0a3c9="" yt:z:up="" srr098401.114049150="" 129="" chr8="" 73321970="" 40="" 24m="" chr11="" 101643970="" 0="" gtgttggtggtttgttttgttgtg="" dgd<gdda7cegeeefeff4=""><;@ AS:i:-4 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:21T2 YT:Z:UP SRR098401.114049247 89 chr9 41024476 0 45M = 41024476 0 GGGCGGTGCGGGGGTGTGAGGCTTTTGGCCAGCATACAGAGACAC BDD8DDHFGHEEEEEAFFEDFFHHHHHHHHHHHHHHHHHHHHHHH AS:i:-20 XN:i:0 XM:i:4 XO:i:0XG:i:0 NM:i:4 MD:Z:2T1T2T7T29 YT:Z:UP SRR098401.114049247 165 chr9 41024476 0 * = 41024476 0 TTGTATTTTGTTGGTTTTGGTTGG CC,/&CAA@@GIG7IDDDDD1CC6 YT:Z:UP SRR098401.114049279 83 chr4 123028176 0 37M = 123028112 -101 TAGGTTGCAGGTATCCTGGTCAGATATAGGAGGACTG A>B@@BABDECEFEHGGHHHHHGHFHGHGHHGHHHGH AS:i:-15 XN:i:0 XM:i:3 XO:i:0 XG:i:0 NM:i:3MD:Z:2T0T2T30 YS:i:-16 YT:Z:CP SRR098401.114049279 163 chr4 123028112 0 34M = 123028176 101 TTTGTTTTTTTTTTTTTTTTGGTTGTGTGTTTGT D6DDCBBBBB4FFFFCCBAB-BCA,?3?=;AAA< AS:i:-16 XS:i:-10 XN:i:0 XM:i:5 XO:i:0XG:i:0 NM:i:5 MD:Z:1A8A9A3C1C7 YS:i:-15 YT:Z:CP SRR098401.114049304 99 chr15 89496360 24 25M = 89496459 121 CTTGGGTAGCGATAGTAGAATTCGT ><<>9:@BA,EEF=FC@BE8B<@CB AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:25 YS:i:-10 YT:Z:CP SRR098401.114049304 147 chr15 89496459 24 22M = 89496360 -121 AACACCCCAAAAAAAACCACCA ?>@9@CC@'@@2A'CB+>>>>@ AS:i:-10 XN:i:0 XM:i:4 XO:i:0 XG:i:0 NM:i:4 MD:Z:8G2G1G2T5 YS:i:0YT:Z:CP SRR098401.114049405 77 * 0 0 * * 0 0 AATGAGCTGAGGCCAGGGCCGCTCCCGGCTGC GHHHEHGFHEGHHHGGFGFECB@CC;><>:=A YT:Z:UP SRR098401.114049405 141 * 0 0 * * 0 0 TCTGTTTTGGTTGGTCTGGTTT @,00@=;:%4>=<1=,0@@<6@ YT:Z:UP SRR098401.114049415 137 chr3 126560201 7 24M = 126560201 0 TGTTGGTTTTTTTTTTTTGTGTTT C,CCCC>CBC:FFFF<,9*@:8;; AS:i:-2 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:18T5 YT:Z:UP SRR098401.114049415 69 chr3 126560201 0 * = 126560201 0 AAAAGGAAATCTCCACAACCGTGCTCCCCCCGGTTCTTGTTTTCGTTG GGGGGFBGBGGFGGGEEEE>80653CACB>B=?A?DDBEA9><9:;>@ YT:Z:UP SRR098401.114049439 153 chr5 131847986 6 21M = 131847986 0 CAAAACCACAACAAAACCAAA @EGE9EB?@<@-@-6@651@@ AS:i:-3 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:13T7 YT:Z:UP SRR098401.114049439 101 chr5 131847986 0 * = 131847986 0 GCCACTGTCCTCCAAAACCCCGCCTGGTCGCTCCCCCGCCCGCTT HGHDGHHHHHHHGHHHECH@BE:2@C@>>B9<<:04?B;B9?BA? YT:Z:UP SRR098401.114049460 73 chr4 28211665 0 20M = 28211665 0 CCCCACCATCTAACCCAGTT FHHHHGHEGGGFFEGGFGGG AS:i:-5 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:14A5 YT:Z:UP SRR098401.114049460 133 chr4 28211665 0 * = 28211665 0 TGTTGTGTGTTTGGGGTGTTTGGTGGTTGG CA11B@C@AC9>2?3B=@79@-@;-@,=@< YT:Z:UP SRR098401.114049495 77 * 0 0 * * 0 0 GCATATAGTAAATACCCTATCCCTCCCTCTGGCCTCCCCC HHHHHHHHHGHHHHHGEGGGCCFFDEEFDFHFEEHFGFGE YT:Z:UP SRR098401.114049495 141 * 0 0 * * 0 0 GGGTTTGGTTGGTGGTTGGGTTTGTTTGTTTGTTT C6C0CCCC6C@84/1@@@@@HHHHHE@%GE?BC?? YT:Z:UP SRR098401.114049544 137 chr5 158711659 1 20M = 158711659 0 GGGTTTTTTTGTTTGCTTTT ?29.??.>@@%@%@@,>@?> AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:20 YT:Z:UP SRR098401.114049544 69 chr5 158711659 0 * = 158711659 0 TATAGGAAATCAATGTGTTGCCCGTTGTCTTCGTCCCCCCCG HHHHGHHHHHHHFHHFGGGG@DADD>>//>6-9<7CFFG?=? YT:Z:UP SRR098401.114049553 89 chr3 196238676 42 25M = 196238676 0 AGGGAGGAAACATTAGTCTTCTTCA B8@4C?GFGGGGGFGGGGGEGGFGG AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:25 YT:Z:UP SRR098401.114049553 165 chr3 196238676 0 * = 196238676 0 TGGTTTGTTGTTGTGTTTGGGTTTGGTGTTGGGGT @6,@>?<>1<1ABA15@>1@C?CB2GDGEGEEGEC YT:Z:UP SRR098401.114049628 153 chr6 4420767 7 21M = 4420767 0 CACCAAAACCAAACCAAACAA BCDDDG6<@@@1D1BB<gggg as:i:-3="" xn:i:0="" xm:i:1="" xo:i:0="" xg:i:0="" nm:i:1="" md:z:13a7="" yt:z:up="" srr098401.114049628="" 101="" chr6="" 4420767="" 0="" *="4420767" 0="" ttaaatatttcatttcccttgttttctttttgccgcc="" hhhhhhhhhhhhhhhhdfhhghhhh6="">@<':@<<

SRR098401.114050028 163 chr16 88249434 0 35M = 88249587 175 TTTCTTTTTTTTTTTTTTTTTGGGTTGGGGTTTTG GGG(GCC%CCFFFFFHHHHHBB8G6;=;)?C7CBC AS:i:-17 XS:i:-12 XN:i:0 XM:i:5 XO:i:0XG:i:0 NM:i:5 MD:Z:3T18A5A0A1C3 YS:i:-5 YT:Z:CP SRR098401.114051567 97 chr2 172534392 1 20M chr4 48927903 0 CTGGGTGGGGGGGGGGGGGG >>>>*=>;;;0DDCD@11@; AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:20 YT:Z:UP SRR098401.114051567 145 chr4 48927903 1 20M chr2 172534392 0 ACAAAACAAAACCCCCCCAC @9<@@@71CC+<><<cc:6b as:i:0="" xn:i:0="" xm:i:0="" xo:i:0="" xg:i:0="" nm:i:0="" md:z:20="" yt:z:up<="" p="">

alignment • 2.7k views
ADD COMMENT
3
Entering edit mode
6.4 years ago
Seigfried ▴ 80

The difference between -S and -s

-s/--skip <int> Skip (i.e. do not align) the first <int> reads or pairs in the input.

-S <sam> File to write SAM alignments to. By default, alignments are written to the "standard out" or "stdout" filehandle (i.e. the console).

ADD COMMENT
1
Entering edit mode
6.4 years ago
MSM55 ▴ 160

There is small mistake in your command, you should use "-S" instead of "-s".

ADD COMMENT

Login before adding your answer.

Traffic: 2580 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6