6/11/2023 0 Comments Nucleobytes 4peaksYou will need to do this for all of the COI sequences that you generated. Trim each forward and reverse sequence based on the presence of Ns (unknown characters). The main change that you will make is adding and subtracting "#" at the beginning of specific lines in order to run each of the individual steps to process the sequences. You will make changes to the sbatch script anytime that you want to run a different command below. #revseq -sequence sequence1-merged.fa -outseq sequence1-merged-rev.faĤ. Here is the command to reverse compliment your merged sequence. # If your top blast hit to the reference inicates that the match is "Strand=PLUS/MINUS" and not "Strand=PLUS/PLUS", you will need to reverse compliment the sequence prior to building a tree. #blastn -db /work/jennifer.bowen/EEMB1105/EXAMPLE-DATA/cap-test/FDA-RSSL.fa -query sequence1-merged.fa -out sequence1-merged-blastout # To blast a sequence against the reference database of COI fish sequences provied by the Ocean Genome Legacy. #merger -asequence sequence1F-trimmed-rev.fa -bsequence sequence1R-trimmed.fa -outfile sequence1-merged.aln -outseq sequence1-merged.fa # Merge the forward and reverse sequences (step 11 of the tutorial) #revseq -sequence sequence1F-trimmed.fa -outseq sequence1F-trimmed-rev.fa # Reverse compliment the forward sequence (*step 10 of the tutorial) #trimseq -sequence *sequence1R.ab1* -outseq *sequence1R-trimmed.fa* -window 20 -percent 5 #trimseq -sequence *sequence1F.ab1* -outseq *sequence1F-trimmed.fa* -window 20 -percent 5 # Trim each forward and reverse sequences (step 9 of the tutorial) I use "#" before any notes I want to remember about this job submission and a "#" in front of commands that I don't want to run. # anything with a "#" in from of it will not be read by the head node except for the "#SBATCH" details above. # You will never need to change any of the details above this line. Just to be sure that you are in the correct location, change to the directory you created that contains all of your COI sequences. You will use "emacs" to create a "sbatch script" identical to the one above in your /scratch/ user.name/FISH-COI/ directory according to the following steps. Here is an example of what one looks like with annotation of what each line means. We can communitcate all of these details in a "Slurm script" aka "bash script" aka "sbatch script". Running commands that process your sequences from the head node is BAD!! The head node is already working hard for other people to distribute the commands that they are submitting, so slowing it down with heavier tasks is very inconsiderate! To "submit a job" aka "run a script", aka "execute a command" we need to create a file that communicates details to the head node about a job we want to submit, such as how much memory we think it will take, and how long we expect the job to run etc. When you log into discovery, you are working from a computer (head node) that controls a huge number of other high performance computers (nodes). MUST READ TO UNDERSTAND THE REST OF THE TUTORIAL: Now that you are all set up with your data and the software that you need to be sequencing genious, you need to know how to run a program on the discovery cluster.
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