Fork this repo - you should do all your work in your fork of this repository.

Write up a strategy for writing a Reference Based PCR Duplicate Removal tool. That is, given a sam file of uniquely mapped reads, remove all PCR duplicates (retain only a single copy of each read). Develop a strategy that avoids loading everything into memory. You should not write any code for this portion of the assignment. Be sure to:

• Define the problem
• Write examples:
  • Include a properly formated input sam file
  • Include a properly formated expected output sam file
• Develop your algorithm using pseudocode
• Determine high level functions
  • Description
  • Function headers
  • Test examples (for individual functions)
  • Return statement

For this portion of the assignment, you should design your algorithm for single-end data, with 96 UMIs. UMI information will be in the QNAME, like so: NS500451:154:HWKTMBGXX:1:11101:15364:1139:GAACAGGT. Discard any UMIs with errors (or error correct, if you're feeling ambitious).

An important part of writing code is reviewing code - both your own and other's. In this portion of the assignment, you will be assigned 3 students' algorithms to review. Be sure to evaluate the following points:

• Does the proposed algorithm make sense to you? Can you follow the logic?
• Does the algorithm do everything it's supposed to do? (see part 1)
• Are proposed functions reasonable? Are they "standalone" pieces of code?

You can find your assigned reviewees on Canvas. You can find your fellow students' repositories at

github.com/<user>/Deduper

Be sure to leave comments on their repositories by creating issues or by commenting on the pull request.

Write your deduper function!

Given a SAM file of uniquely mapped reads, remove all PCR duplicates (retain only a single copy of each read). Remember:

• Samtools sort
• Adjust for soft clipping
• Single-end reads
• Known UMIs
• Considerations:
  • Millions of reads – avoid loading everything into memory!
  • Be sure to utilize functions appropriately
  • Appropriately comment code and include doc strings
• CHALLENGE: Include options for
  • Single-end vs paired-end
  • Known UMIs vs randomers
  • Choice of duplicate written to file

You MUST:

• Write Python 3 compatible code
• Include the following argparse options
  • -f, --file: required arg, absolute file path
  • -p, --paired: optional arg, designates file is paired end (not single-end)
  • -u, --umi: optional arg, designates file containing the list of UMIs (unset if randomers instead of UMIs)
  • -h, --help: optional arg, prints a USEFUL help message (see argparse docs)
    • If your script is not capable of dealing with a particular option (ex: no paired-end functionality), your script should print an error message and quit
• Output the first read encountered if duplicates are found
  • You may include an additional argument to designate output of a different read (highest quality or random or ???)
• Output a properly formatted SAM file with “_deduped” appended to the filename
• Name your python script <your_last_name>_deduper.py