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@@ -15,7 +15,7 @@ Bowtie Getting Started Guide |
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genome. To use Bowtie to align those reads, issue the following
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command. If you get an error message "command not found", try adding
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a "./" before the "bowtie".
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-
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+
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bowtie e_coli reads/e_coli_1000.fq
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The first argument to bowtie is the basename of the index for the
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@@ -26,7 +26,7 @@ Bowtie Getting Started Guide |
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about a minute. You will see bowtie print many lines of output. Each
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line is an alignment for a read. The name of the aligned read appears
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in the leftmost column. The final line should say "Reported 698
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- alignments to 1 output stream(s)" or something similar.
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+ alignments to 1 output stream(s)" or something similar.
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Next, issue this command:
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@@ -82,14 +82,15 @@ Bowtie Getting Started Guide |
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The pre-built E. coli index included with Bowtie is built from the
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sequence for strain 536, known to cause urinary tract infections. We
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will create a new index from the sequence of E. coli strain O157:H7, a
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- strain known to cause food poisoning. Download the sequence file from:
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+ strain known to cause food poisoning. Download and decompress the
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+ sequence file from:
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- ftp://ftp.ncbi.nlm.nih.gov/genomes/Bacteria/Escherichia_coli_O157H7/NC_002127.fna
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+ ftp://ftp.ncbi.nlm.nih.gov/genomes/refseq/bacteria/Escherichia_coli/all_assembly_versions/GCF_000513035.1_E._coli_O157/GCF_000513035.1_E._coli_O157_genomic.fna.gz
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- When the sequence file is finished downloading, move it to the Bowtie
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+ Once it has been downloaded and decompressed, move it to the Bowtie
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install directory and issue this command:
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- bowtie-build NC_002127.fna e_coli_O157_H7
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+ bowtie-build GCF_000513035.1_E._coli_O157_genomic.fna e_coli_O157_H7
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The command should finish quickly, and print several lines of status
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messages. When the command has completed, note that the current
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@@ -100,7 +101,7 @@ Bowtie Getting Started Guide |
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To test that the index is properly installed, issue this command:
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- bowtie -c e_coli_O157_H7 GCGTGAGCTATGAGAAAGCGCCACGCTTCC
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+ bowtie -c e_coli_O157_H7 GAACCGTATTCACCCGCCATCCCCATGCCG
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If the index is installed properly, this command should print a single
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alignment and then exit.
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@@ -132,16 +133,16 @@ Bowtie Getting Started Guide |
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accessible in the PATH.
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samtools view -bS -o ec_snp.bam ec_snp.sam
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-
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+
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Next, we sort the BAM file, in preparation for SNP calling:
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samtools sort ec_snp.bam ec_snp.sorted
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-
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+
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We now have a sorted BAM file called ec_snp.sorted.bam. Sorted BAM is
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a useful format because the alignments are both compressed, which is
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convenient for long-term storage, and sorted, which is conveneint for
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variant discovery. Finally, we call variants from the Sorted BAM:
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-
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+
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samtools pileup -cv -f genomes/NC_008253.fna ec_snp.sorted.bam
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For this sample data, the 'samtools pileup' command should print
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