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Homework #4
Due April 12th, 11:59pm
Each homework submission must include:
• An archive (.zip or .gz) file of the source code containing:
o The makefile used to compile the code on Monsoon (5pts) o All .cpp and .h files (5pts)
• A full write-up (.pdf of .doc) file containing answers to homework’s questions (5pts), including the exact command line needed to execute every subproblem of the homework
The source code must follow the following guidelines:
• No external libraries that implement data structures discussed in class are allowed, unless specifically stated as part of the problem definition. Standard input/output and utilities libraries (e.g. math.h) are ok.
• All external data sources (e.g. input data) must be passed in as a command line argument (no hardcoded paths within the source code (5pts).
• Solutions to sub-problems must be executable separately from each other. For example, via a special flag passed as command line argument (5pts)
For this homework, you will need to use the sequence fragments you used as part of Homework #1 and #2, located at /common/contrib/classroom/inf503/hw_dataset.fa . You will also need the genome sequence for B. anthracis bacterium located at: common/contrib/classroom/inf503/test_genome.fasta
• This genome file contains a header (denoted by ‘ ’) followed by ~5.2 million characters of its genomic code (alphabet A, C, G, T)
• Please be aware that the genome is spread across multiple lines of the file (see insert)
Problem #1 (of 1): Having a BLAST
Create a class called BLAST_DB, which would facilitate seed-based searching. Use the hash table datastructure with chaining collision resolution as the primary data structure in the class. The class must be able to take a genome sequence, break it down into all possible seeds with a given word size (for this homework, word size = 11), and store these seeds (along with their location in the genome) in the hash table. Use Radix / division scheme for hash function implementation. Seed searching should be done via the hash table, seed extension should be done using a Needleman-Wunsch algorithm. Assume a gap penalty of -1, mismatch penalty of -1, and match score of +2.
At minimum, the class must contain (25pts):
• A constructor
• A destructor
• A function to disassemble the genome into seeds of a given size and store them into the hash table data structure.
• A function to implement a Needleman Wunsch algorithm.
• A function to search a given query withing the BLAST_DB class. Your function must return the genome location of the best hit and score for the best hit.
A. (50 pts) Basic seed-based searching. Implement seed-based Needleman-Wunsch searching algorithm. For this you will need to (a) precompute and store all possible 11-mer seeds of the B. anthracis genome in the BLAST_DB’s hash-table data structure – use m = 10,000,000 to ensure good performance and do not forget to store seed’s location in the genome; (b) disassemble each query into all possible 11-mers, storing the location of each 11-mer in the original query; (c) search the query’s 11-mers in the seeds stored in BLAST_DB and store each seed-hit in a stack or queue data structure (d) expand each seed into a full alignment by selecting a proper portion of the genome sequence and calling your Needleman-Wunsch function; and (e) compare scores for all seed extensions to find the best alignment.
• Randomly select 10K, 100K, and 1M (million) 50-mer fragments from the B. anthracis genome and use these fragments to query the BLAST_DB containing B. anthracis. How many fragments were you able to find and how long did it take?
• Randomly select 10K, 100K, and 1M (million) 50-mer fragments from the B. anthracis genome and introduce a 5% per-base error rate (every character has a 5% change of being changed to some other random character). Use these error-filled fragments to query the BLAST_DB containing B. anthracis. How many fragments were you able to find?
• Read the High Throughput Sequence reads dataset you used for homework #1 and #2 (located at /common/contrib/classroom/inf503/hw_dataset.fa). For this assignment, you can completely disregard the headers of the sequence fragments (i.e. R0_0_1…). Search the entire contents of this dataset in the BLAST_DB. How many perfect hits did you find? (hint: perfect hit’s score = 100) Please note that depending on the efficiency of your algorithm, this step may take a long time. First estimate the total time using 1,000, 10,000, and 100,000 queries – if total time estimate is greater than 24 CPU hours, provide estimate rather than exact number.
B. (15 points extra credit) Making it pretty. Adapt your Needleman-Wunsch function to also return a human-friendly alignment (see below). You will need to actually perform a traceback and identify matching characters and gaps (if any). Hint: use a struct of 3 character arrays, 2 for sequences one for alignment codes {x, I , } – use whitespace for gaps).
GATTA
| x||
G_CTA



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