## [Bowtie](http://bowtie-bio.sourceforge.net/) align reads on _indexed_ genomes {width="600"} ## Preliminary Note :warning: For the following steps, you will need 2 programs which an admin (with admin rights) has already installed - system-wide - for you, using the following command: ``` sudo apt update && apt install -y bowtie samtools ``` ## Prepare dmel_r6.54 bowtie index (*Drosophila* genome) !!! note "Bowtie indexing command line" ``` bowtie-build --threads 6 dmel-all-chromosome-r6.54.fasta dmel.r6.54 ``` :clock230: This step should take about 2-3 min ## Align the clipped fasta reads to dmel.r6.54 using `bowtie` ``` bowtie dmel.r6.54 -f clipped_GRH-103_R1.fasta \ -v 0 \ -k 1 \ -p 3 \ --al dmel_matched_GRH-103.fa \ --un unmatched_GRH-103.fa \ -S > GRH-103.sam ``` :clock230: This step should take about 1 min ??? info "The bowtie alignment command explained" - `bowtie dmel.r6.54 -f clipped_GRH-103_R1.fasta` # tells bowtie where is the index and the input clipped_GRH-103_R1.fasta - `-v 0 -k 1 -p 3` # These are bowtie options - `--al dmel_matched_GRH-103.fa` # aligned reads will be in the dmel_matched_GRH-103.fa file - `--un unmatched_GRH-103.fa` # Unaligned reads will be in the unmatched_GRH-103.fa file - `-S > GRH-103.sam` # tells bowtie to return an alignement file in SAM format (-S) -S > GRH-103.sam ## Convert SAM file to BAM file and sort the alignments by chromosome positions ``` samtools view -Sb -@ 3 GRH-103.sam | samtools sort -@ 3 -o GRH-103.bam ``` Check the result using ``` samtools view GRH-103.bam | more ```