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MacManes_MapQuest.rst

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Map Quest

I am going to give you some data and a genome (well, 500Mb of the genome), I want you to quality trim and map using the Trimming lecture and the Mapping Lecture.

** Launch an AMI. For this exercise, try a c4.4xlarge with a 200Gb EBS volume.

You will need to install skewer, bwa, and samtools. Maybe try brew???

Download the genome and the reads

https://s3.amazonaws.com/macmanes_share/Cavia.fa
curl -LO ftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR636/SRR636898/SRR636898_1.fastq.gz
curl -LO ftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR636/SRR636898/SRR636898_2.fastq.gz

Use Skewer to trim reads

  1. For people whose 1st name starts with between A-F, skip this step to allow us to evaluate the performance of mapping untrimmed reads.
  2. Everybody else, choose a random number between 1 and 30 and trim at that level..

Map the reads with BWA You will need to figure out how to map paired end reads. May luck be ever in your favor.. Also, there is google and the help info.

Type bwa mem to get a list of the options. You will need to index the genome using bwa index before you map.

bwa mem [you need to figure out the command] \
| samtools view -@10 -Sb - \
| samtools sort -T sort -O bam -@10 -o sorted.bam -

Evaluate your mapping data

samtools flagstat sorted.bam

Add your data about mappability

https://docs.google.com/spreadsheets/d/1svCJMefIy-BTBSWl5Vixf0UPgLIGOrp5u6YRoyMbSng/edit?usp=sharing

Super bonus points to anybody that maps with a differnt mapper e.g., Bowtie, STAR, VelociMapper, HiSat2, etc..

TERMINATE YOUR INSTANCE!!!