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fixing e coli link and instructions
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BenLangmead committed Nov 8, 2016
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21 changes: 11 additions & 10 deletions TUTORIAL
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Expand Up @@ -15,7 +15,7 @@ Bowtie Getting Started Guide
genome. To use Bowtie to align those reads, issue the following
command. If you get an error message "command not found", try adding
a "./" before the "bowtie".

bowtie e_coli reads/e_coli_1000.fq

The first argument to bowtie is the basename of the index for the
Expand All @@ -26,7 +26,7 @@ Bowtie Getting Started Guide
about a minute. You will see bowtie print many lines of output. Each
line is an alignment for a read. The name of the aligned read appears
in the leftmost column. The final line should say "Reported 698
alignments to 1 output stream(s)" or something similar.
alignments to 1 output stream(s)" or something similar.

Next, issue this command:

Expand Down Expand Up @@ -82,14 +82,15 @@ Bowtie Getting Started Guide
The pre-built E. coli index included with Bowtie is built from the
sequence for strain 536, known to cause urinary tract infections. We
will create a new index from the sequence of E. coli strain O157:H7, a
strain known to cause food poisoning. Download the sequence file from:
strain known to cause food poisoning. Download and decompress the
sequence file from:

ftp://ftp.ncbi.nlm.nih.gov/genomes/Bacteria/Escherichia_coli_O157H7/NC_002127.fna
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

When the sequence file is finished downloading, move it to the Bowtie
Once it has been downloaded and decompressed, move it to the Bowtie
install directory and issue this command:

bowtie-build NC_002127.fna e_coli_O157_H7
bowtie-build GCF_000513035.1_E._coli_O157_genomic.fna e_coli_O157_H7

The command should finish quickly, and print several lines of status
messages. When the command has completed, note that the current
Expand All @@ -100,7 +101,7 @@ Bowtie Getting Started Guide

To test that the index is properly installed, issue this command:

bowtie -c e_coli_O157_H7 GCGTGAGCTATGAGAAAGCGCCACGCTTCC
bowtie -c e_coli_O157_H7 GAACCGTATTCACCCGCCATCCCCATGCCG

If the index is installed properly, this command should print a single
alignment and then exit.
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accessible in the PATH.

samtools view -bS -o ec_snp.bam ec_snp.sam

Next, we sort the BAM file, in preparation for SNP calling:

samtools sort ec_snp.bam ec_snp.sorted

We now have a sorted BAM file called ec_snp.sorted.bam. Sorted BAM is
a useful format because the alignments are both compressed, which is
convenient for long-term storage, and sorted, which is conveneint for
variant discovery. Finally, we call variants from the Sorted BAM:

samtools pileup -cv -f genomes/NC_008253.fna ec_snp.sorted.bam

For this sample data, the 'samtools pileup' command should print
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