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(Solved) : Bioinformatics Need Use Galaxy Set Experiment Given Files Unsure Begin Distributing Galaxy Q35533032 . . .

BIOINFORMATICS

I need to use Galaxy to set up an experiment on some givenfiles, but I am unsure how to begin.

  1. I am distributing a Galaxy History called:CG19FinalProjectDNADatasets containing:
    • Sacce1_AssemblyScaffolds.fasta <- Saccharomycescerevisiae Wild-Type Genome
    • Sacce1.ExternalModels.gff3 <- Saccharomycescerevisiae Wild-Type Genome Annotations
    • Scerevisiae_RAMutant_Genome.v02.fa <- Saccharomycescerevisiae Mutant Genome
    • Sacce1_chrVI-VII-VIII.fasta <- Saccharomycescerevisiae Chromosomes VI/VII/VIII Wild-Type Genome
    • All Datasets are from the Wild-TypeGenome
    • Dataset01:
      • SRR352492.sra_1.fastq
      • SRR352492.sra_2.fastq
    • Dataset02:
      • SRR352384.sra_1.fastq
      • SRR352384.sra_2.fastq
    • Dataset03:
      • SRR452441.csra_1.fastq
      • SRR452441.csra_2.fastq
    • The Datasets have already been Quality Controlled
    • Saccharomyces cerevisiae Wild-Type ChromosomesVI/VII/VIII is being provided for you to test/optimize your mappingconditions
  2. Your task is:
    • To use these NGS Datasets to identify any polymorphisms thatmight be present in theSaccharomyces cerevisiae MutantGenome
    • You are welcome to validate your findings using other methods(e.g., alignments, Blast, etc), but your primary conclusionsmust be based on NGS Reads mapping
    • Your findings must also be validated by IGVfigures displaying the presence/absence of the polymorphisms youhave identified. In other words, each polymorphism claim must besupported by an IGV figure
    • All your findings must be properlydocumented
    • A Final Project Report and FinalProject Presentation must be prepared according to theinstructions outlined below
  3. Recommendations:
    • Start by looking at your data, study the data
    • Design a logical and simple Experimental Strategy. Rememberthere is no a single way to address this problem
    • Learn how to use and try to understand the following tools inGalaxy:
      • Bowtie map reads against reference genome (output = unsortedsam file)
      • Bowtie2 – map reads against reference genome (output = sortedbam file)
      • After you settle on Bowtie and/or Bowtie2, do not change thisparameter
      • Pay special attention to the options provided to generate filescontaining aligned and/or unaligned reads. Remember, you might needto use those parameters to demonstrate your assumptions or testyour hypothesis
      • sort a BAM file <= Sorting is imperative after theBam file is generated and before other processing canoccur
      • Flagstat tabulate descriptive stats for BAM dataset
      • BAM mapping statistics samtools idxstats
      • Filter BAM datasets on a variety of attributes
      • BAM-to-SAM convert BAM to SAM and SAM-to-BAM convert SAM to BAM(when needed)
    • I suggest using the test chromosome you must decide on whichone of these tools is best for you task
    • Display the data on IGV
      • When using BAM files, remember to download, in addition to theBAM file, its accompanying BAM_INDEX file
    • If you need to assemble reads, let me know, and I will show youhow to do it
    • Include Controls

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