*.bam *.bai *.fai *.glf* *.pileup* calDepth ex1*.rg ex1.bcf ex1.vcf >seq1 CACTAGTGGCTCATTGTAAATGTGTGGTTTAACTCGTCCATGGCCCAGCATTAGGGAGCT GTGGACCCTGCAGCCTGGCTGTGGGGGCCGCAGTGGCTGAGGGGTGCAGAGCCGAGTCAC GGGGTTGCCAGCACAGGGGCTTAACCTCTGGTGACTGCCAGAGCTGCTGGCAAGCTAGAG TCCCATTTGGAGCCCCTCTAAGCCGTTCTATTTGTAATGAAAACTATATTTATGCTATTC AGTTCTAAATATAGAAATTGAAACAGCTGTGTTTAGTGCCTTTGTTCAACCCCCTTGCAA CAACCTTGAGAACCCCAGGGAATTTGTCAATGTCAGGGAAGGAGCATTTTGTCAGTTACC AAATGTGTTTATTACCAGAGGGATGGAGGGAAGAGGGACGCTGAAGAACTTTGATGCCCT CTTCTTCCAAAGATGAAACGCGTAACTGCGCTCTCATTCACTCCAGCTCCCTGTCACCCA ATGGACCTGTGATATCTGGATTCTGGGAAATTCTTCATCCTGGACCCTGAGAGATTCTGC AGCCCAGCTCCAGATTGCTTGTGGTCTGACAGGCTGCAACTGTGAGCCATCACAATGAAC AACAGGAAGAAAAGGTCTTTCAAAAGGTGATGTGTGTTCTCATCAACCTCATACACACAC ATGGTTTAGGGGTATAATACCTCTACATGGCTGATTATGAAAACAATGTTCCCCAGATAC CATCCCTGTCTTACTTCCAGCTCCCCAGAGGGAAAGCTTTCAACGCTTCTAGCCATTTCT TTTGGCATTTGCCTTCAGACCCTACACGAATGCGTCTCTACCACAGGGGGCTGCGCGGTT TCCCATCATGAAGCACTGAACTTCCACGTCTCATCTAGGGGAACAGGGAGGTGCACTAAT GCGCTCCACGCCCAAGCCCTTCTCACAGTTTCTGCCCCCAGCATGGTTGTACTGGGCAAT ACATGAGATTATTAGGAAATGCTTTACTGTCATAACTATGAAGAGACTATTGCCAGATGA ACCACACATTAATACTATGTTTCTTATCTGCACATTACTACCCTGCAATTAATATAATTG TGTCCATGTACACACGCTGTCCTATGTACTTATCATGACTCTATCCCAAATTCCCAATTA CGTCCTATCTTCTTCTTAGGGAAGAACAGCTTAGGTATCAATTTGGTGTTCTGTGTAAAG TCTCAGGGAGCCGTCCGTGTCCTCCCATCTGGCCTCGTCCACACTGGTTCTCTTGAAAGC TTGGGCTGTAATGATGCCCCTTGGCCATCACCCAGTCCCTGCCCCATCTCTTGTAATCTC TCTCCTTTTTGCTGCATCCCTGTCTTCCTCTGTCTTGATTTACTTGTTGTTGGTTTTCTG TTTCTTTGTTTGATTTGGTGGAAGACATAATCCCACGCTTCCTATGGAAAGGTTGTTGGG AGATTTTTAATGATTCCTCAATGTTAAAATGTCTATTTTTGTCTTGACACCCAACTAATA TTTGTCTGAGCAAAACAGTCTAGATGAGAGAGAACTTCCCTGGAGGTCTGATGGCGTTTC TCCCTCGTCTTCTTA >seq2 TTCAAATGAACTTCTGTAATTGAAAAATTCATTTAAGAAATTACAAAATATAGTTGAAAG CTCTAACAATAGACTAAACCAAGCAGAAGAAAGAGGTTCAGAACTTGAAGACAAGTCTCT TATGAATTAACCCAGTCAGACAAAAATAAAGAAAAAAATTTTAAAAATGAACAGAGCTTT CAAGAAGTATGAGATTATGTAAAGTAACTGAACCTATGAGTCACAGGTATTCCTGAGGAA AAAGAAAAAGTGAGAAGTTTGGAAAAACTATTTGAGGAAGTAATTGGGGAAAACCTCTTT AGTCTTGCTAGAGATTTAGACATCTAAATGAAAGAGGCTCAAAGAATGCCAGGAAGATAC ATTGCAAGACAGACTTCATCAAGATATGTAGTCATCAGACTATCTAAAGTCAACATGAAG GAAAAAAATTCTAAAATCAGCAAGAGAAAAGCATACAGTCATCTATAAAGGAAATCCCAT CAGAATAACAATGGGCTTCTCAGCAGAAACCTTACAAGCCAGAAGAGATTGGATCTAATT TTTGGACTTCTTAAAGAAAAAAAAACCTGTCAAACACGAATGTTATGCCCTGCTAAACTA AGCATCATAAATGAAGGGGAAATAAAGTCAAGTCTTTCCTGACAAGCAAATGCTAAGATA ATTCATCATCACTAAACCAGTCCTATAAGAAATGCTCAAAAGAATTGTAAAAGTCAAAAT TAAAGTTCAATACTCACCATCATAAATACACACAAAAGTACAAAACTCACAGGTTTTATA AAACAATTGAGACTACAGAGCAACTAGGTAAAAAATTAACATTACAACAGGAACAAAACC TCATATATCAATATTAACTTTGAATAAAAAGGGATTAAATTCCCCCACTTAAGAGATATA GATTGGCAGAACAGATTTAAAAACATGAACTAACTATATGCTGTTTACAAGAAACTCATT AATAAAGACATGAGTTCAGGTAAAGGGGTGGAAAAAGATGTTCTACGCAAACAGAAACCA AATGAGAGAAGGAGTAGCTATACTTATATCAGATAAAGCACACTTTAAATCAACAACAGT AAAATAAAACAAAGGAGGTCATCATACAATGATAAAAAGATCAATTCAGCAAGAAGATAT AACCATCCTACTAAATACATATGCACCTAACACAAGACTACCCAGATTCATAAAACAAAT ACTACTAGACCTAAGAGGGATGAGAAATTACCTAATTGGTACAATGTACAATATTCTGAT GATGGTTACACTAAAAGCCCATACTTTACTGCTACTCAATATATCCATGTAACAAATCTG CGCTTGTACTTCTAAATCTATAAAAAAATTAAAATTTAACAAAAGTAAATAAAACACATA GCTAAAACTAAAAAAGCAAAAACAAAAACTATGCTAAGTATTGGTAAAGATGTGGGGAAA AAAGTAAACTCTCAAATATTGCTAGTGGGAGTATAAATTGTTTTCCACTTTGGAAAACAA TTTGGTAATTTCGTTTTTTTTTTTTTCTTTTCTCTTTTTTTTTTTTTTTTTTTTGCATGC CAGAAAAAAATATTTACAGTAACT I\ɎJv]ѩL%,lKёl?g^2qItyJ8Nj5cX?dBF yChuKL1xs=?wmAhuA8>P7n{\e)gƲsc M! pB<9SUIH.D>L B"[t{+`̠ROmeDGQ>ݶu HzQp:p2Jim<9zgֲ {H~Rײ?LY 0_j\LE#0*BR)D{)LnGXm ⟅Zz9r1 W1W-ZSpQhHD^Vg }ç cŻⵆߩ/r4$ks8zfr1JFIb:3^EMtPUKr1l7"Y5!EcL2O~16#,b48PR0SisA ފ^K5Vgw>rR%au Vπ2 Q0rwWz 'f +[rDQltw@dL_Dr .^dgݳ ῀jf'n q'B7;%z x=ĴibL>!xz,/QG@pzQ1vD(-^kOФv.dyj]WR6m^9aj7uHsj2Jls5Rzt[=)LJwJWd2 i%HAشQv%D`DU2#R,-z6oHIj T=-O;}[VS&mk?~ӽ+w_^_uߜP0}Fێ| utB:$4$/M$t㎍; Ln/7Q{rFy&!K-᫛R=гT3˺3w00S _,ڡhInQm`m5x10PvԠ9hq%w{6W7QN2G.!_糟5)je'0=`j!}0/{){7"9?ϕ6rJ#CJ yL;gPРj!5N1Lnⷷv9J 41!bOx$}DWX-Sp"QJA)iϹ)b:~>?s M@n Ixҋܪx. ־J^L%T mC8n|Xd0'+rr%|rbZXx>iJ=ԓa|Sfv sޜ*fs⤮ :,J7 iy$%DGk*U"o] XE6嵠~dM6$mRayܭvOLPƶpܒtգ.ɒQ/)9QzGˠ299nLE/@H`P_4~PuciUL.* HR6ͧh,6<>M OiOsΙ-šC. 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File ex1.fa contains two sequences cut from the human genome build36. They were extracted with command: samtools faidx human_b36.fa 2:2043966-2045540 20:67967-69550 Sequence names were changed manually for simplicity. File ex1.sam.gz contains MAQ alignments extracted with: (samtools view NA18507_maq.bam 2:2044001-2045500; samtools view NA18507_maq.bam 20:68001-69500) and processed with `samtools fixmate' to make it self-consistent as a standalone alignment. To try samtools, you may run the following commands. Index the reference FASTA. samtools faidx ex1.fa Convert the (headerless) SAM file to BAM. Note if we had used "samtools view -h" above to create the ex1.sam.gz then we could omit the "-t ex1.fa.fai" option here. samtools view -S -b -t ex1.fa.fai -o ex1.bam ex1.sam.gz Build an index for the BAM file: samtools index ex1.bam View a portion of the BAM file: samtools view ex1.bam seq2:450-550 Visually inspect the alignments at the same location: samtools tview -p seq2:450 ex1.bam ex1.fa View the data in pileup format: samtools mpileup -f ex1.fa ex1.bam Generate an uncompressed VCF file of variants: samtools mpileup -vu -f ex1.fa ex1.bam > ex1.vcf Generate a compressed VCF file of variants: samtools mpileup -g -f ex1.fa ex1.bam > ex1.bcf