Week 1
  Monday Tuesday Wednesday Thursday Friday Saturday Sunday Lecture 1 Introduction to course and best laboratory practices - Rapin, Fernando, Hill, Misra
    01-May 02-May 03-May 04-May 05-May 06-May Lecture 2 Units and conversions - Hill
9:00   Lecture 1 How to Lecture 5 Lecture 8     Lecture 3 Pipets and pipeting - Hill
      Search     Transfer plates   Lecture 4 Measurement of nucleic acids - Dumonceaux
10:00   Lecture 2 the Lecture 6 Lab 4 from   Lecture 5 Buffers and pH
      literature   Set up PCR incubator to   Lecture 6 Statistics and Experimental design
11:00   Lecture 3     Begin PCR frig   Lecture 7 Purification of DNA, RNA and Proteins
                Lecture 8 PCR and cloning of PCR products
12:00              
               
1:00   Lab 1 Lecture 4 Lecture 7 Set up gel    
                Lab 1 Problem sets - units and conversiton
2:00     Lab 3 Lab 4       Lab 2 Pipets and pipeting
        Triple prep       Lab 3 Nucleic acid measurments
3:00   Lab 2     Electrophoresis   Transfer Lab 4 Triple Prep purification of DNA, RNA and proteins from the same sample
              colonies PCR
4:00         Clone products   to broth Cloning of PCR products
          into pCRTOPO     Screening of colonies for plasmids with inserts
5:00              
               
Week 2
  Monday Tuesday Wednesday Thursday Friday Saturday Sunday Lecture 9 Cloning of DNA fragments – restriction endonucleases, plasmid vectors, uses
  07-May 08-May 09-May 10-May 11-May 12-May 13-May Lecture 10 Determining DNA sequence
9:00 Lecture 9 Lecture 10 Lecture 11 transfer colonies Lecture 13     Lecture 11,12 Basic and practical informatics – restriction endonuclease sites, sequence databases, mRNA and genomic sequences, introns and exons, comparing sequences, designing primers, degenerate primers
        to broth        
10:00 Lab 4 purify plasmid Lab 5 Lecture 12 Purify DNA     Lecture 13 qRT-PCR
  purify plasmid DNA transform   midiprep    
11:00 DNA by Qiaprep E. coli   Qiagen kit     Lab 5 clone DNA fragment in expression vector, midiprep purification of DNA
  miniprep send for Plate         Lab 6 Searching sequence databases, introns and exons, BLAST etc. CLUSTAL
12:00 restrition digest sequencing          
               
1:00 gel Lab 5 Lab 6        
  electrophoresis restriction digest computer        
2:00     excercises screen colonies      
    gel electro.          
3:00              
    elute          
4:00 set up ligate ON.          
  overnight     transfer to      
5:00 cultures     culture      
               
Week 3
  Monday Tuesday Wednesday Thursday Friday Saturday Sunday Lecture 14 Separation of proteins, protein expression and purification
  14-May 15-May 16-May 17-May 18-May 19-May 20-May Lecture 15 Detection of proteins – immunological – western blots, immunofluorescence
9:00 Lecture 14 Lecture 15 Lecture 16 Lecture 17 Lecture 18     Lecture 16 Ectopic expression of proteins in mammalian cells
                Lecture 17 Site directed mutagenesis
10:00 Lab 7 Lab 8 process blots transform cells screen     Lecture 18 Detection gene expression – surrogate genes, transient transfection
  Set up qRT-PCR Prepare gel process slides   colonies for    
11:00     immunofluor.   mutants     Lab 7 qRT-PCR
                Lab 8 SD-PAGE and immunological detection of proteins
12:00   load gel           Lab 9 site-directed mutagenesis
    electrophoresis          
1:00 Lab 7 fix cells for Lab 9        
  Go over immunofluor. mutagenesis        
2:00 qRT-PCR transfer to set up PCR        
  results membrane          
3:00              
               
4:00   blot overnight          
               
5:00              
               
Week 4
  Monday Tuesday Wednesday Thursday Friday Saturday Sunday Lecture 19 Suppressing gene expression - siRNA
  21-May 22-May 23-May 24-May 25-May 26-May 27-May Lecture 20 TBA
9:00   Lecture 19 Lecture 20 Lecture 21 Lecture 22     Lecture 21 TBA
                Lecture 22 TBA
10:00   Lab 10 Lab 10 Lab 10 Lab 10    
    day 0 day 1 day 2 day 3     Lab 10 Measure growth rate of cells - WST assay
11:00   measurements measurements measurements measurements     Lab 11 Determine in mutation changes phenotype of gene
    treat-           Lab 12 Go over results
12:00   infect (Rapin) Immunofluorescence Harvest cells      
    cultures   CAT and Lac-Z      
1:00   Lab 11   assays Lab 12    
    transfect cells          
2:00              
               
3:00              
               
4:00              
               
5:00              
               
Week 5
  Monday Tuesday Wednesday Thursday Friday Saturday Sunday Lecture 23 Bioplex bead-based hybridization - Dumonceaux
  28-May 29-May 30-May 31-May 01-Jun 02-Jun 03-Jun Lecture 24 Campylobacter - ecology and diagnostics - Chaban
9:00 Lecture 23 Lecture 24 Lecture 25 Lecture 26       Lecture 25 Hybridization probe design - Dumonceaux
                Lecture 26 Vaginal microbiome and women's health
10:00              
          check results     Lab 13 Detection of DNA sequences in complex samples using bead-based hybridization (Bioplex)
11:00         of PCR    
               
12:00         T7 exo    
          treatment    
1:00 Lab 13 plasmid dilution Demo      
  start isolation, series for bead counts Bioplex    
2:00 E. coli quantification, standard   reading    
  cultures for confirm curves fecal DNA data    
3:00 plasmid insert by   extraction collection    
  purification PCR set up SYBR        
4:00   rest. Endo. green set up      
      qPCR PCR      
5:00              
               
Room Lectures
All mornings WCVM 2102 except May 15 WCVM 2103
PM May 2 WCVM 2103
Room Lab