Undergrad Research Experience

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Revision as of 14:45, 17 September 2020 by imported>Weigang (→‎Project 3. Clostridium transcriptome analysis)
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Fall 2020

Participants

  • Eamen Ho: Volunteer research assistant
  • Ramandeep Singh: BIOL 48002
  • Desiree Pante: BIOL 48001
  • Afsana Rahman: Volunteer research assistant
  • Roman Shimonov: BIOL 48002
  • Justin Hiraldo: BIOL 48002
  • Zaheen Hossain: Volunteer research assistant
  • Jerry Sebastian: Volunteer research assistant

Schedule

  • Tuesdays at 12 noon - 2pm by Zoom
  • Sept 1, 2020. Week 1. Meet & Greet; Intro to projects
  • Sept 8, 2020. Week 2. Presentations (background, data, and methods), based on assigned readings

Project 1. Structure & evolution of multipartite genome of Lyme disease bacteria

  • Participants: Desiree & Ramon (Summer 2020), Jerry
  • Readings
  • Data set: lp54 & cp26 plasmids
  • TO DO:
    • Week 1. 9/8/2020, 12 noon: 5-slides presentation on multipartite bacterial genome evolution (based on the paper above)
    • Week 2. 9/15, 12 noon: Use prorgram codonO to calculate codon bias (SCUO) for replicons (n=23) on Borrelia burgdorferi B31 genome
    • Week 3. 9/22, 12 noon: codonO paper presentation (Jerry)

Project 2. OspC Cross-reactivity analysis

  • Participants: Justin, Roman
  • Readings: Ivanova et al (2009)
  • Tool: ImageJ
  • Data set (to be sent)
  • To Do
    • Week 1. 9/8/2020 12 noon: 5-slide presentation on background, material & methods, and data capture using ImageJ
    • Week 2. 9/15: Create Excel sheet to capture immunoblot intensities on C3H mice & P.lucus. Capture background for each serum. Getting ready to makes plots in R/Rstudio

Project 3. Clostridium transcriptome analysis

  • Participants: Eaman, Zaheen
  • Readings
  • Data set: posted on "genometracker.org"
    • Wild type transcriptome at 12 hour, paired-end read files:
    • /home/azureuser/18134XR-29-01_S0_L001_R1_001.fastq.gz
    • /home/azureuser/18134XR-29-01_S0_L001_R2_001.fastq.gz
  • To Do
    • Week 1. 9/8/2020 12 noon:
      • A short presentation on C. diff transcriptome (one of the 2 papers above)
      • Demo on read quality using FastQC and mapping reads to reference genomes with bowtie
    • Week 2. Use HT-Seq to quantify RNA abundance for C. diff genes.
    • Commands

According to: reference; Bowtie website

bioseq -i'genbank' R20291.gb > ref.fa # make FASTA file
bowtie2-build ref.fa index # build index
# -S: sam output (otherwise bam) 
bowtie2 -x index -S 18134XR.sam -1 ../18134XR-29-01_S0_L001_R1_001.fastq.gz -2 ../18134XR-29-01_S0_L001_R2_001.fastq.gz
# ref.gff3: need to run sed "s/Chromosome/FN545816/"
# need to use "-i"; default is "gene_id"
htseq-count -m union --stranded=yes 18134XR-29-01.sam ~/xingmin-cdiff/ref.gff3 -i=Parent > 18134XR-29-01.counts
samtools view -b 18134XR-29-01.sam -o 18134XR-29-01.bam # compress sam file into bam file

Project 4. Protein classification using natural language processing