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First International Molecular Methods in Food Microbiology Symposium and Workshop Series:
March 30th – April 3rd 2009; Mahidol University; Bangkok Thailand
Organized by:
Mahidol University; Bangkok, Thailand
National Research Council of Thailand; Bangkok, Thailand
Colorado State University; Fort Collins, CO, United States
Cornell University; Ithaca, NY, United States
Silliker North America Food Science Center; South Holland, IL, United States
Click here to download the schedule 
Lecture series and symposium schedule:
All Times are Local Bangkok Time (Greenwich Mean Time + 7 hours)
Sunday, March 29th: Arrival in Bangkok
Monday, March 30th (Lecture Series Day 1; TBA):
8:00 a.m. Check-in and Complete Pre-evaluations
8:30 a.m. Welcome and Introduction
8:45 a.m. Fundamentals: Nucleic Acids, DNA Replication, Transcription and Translation and Application to Molecular Detection Assay Development (Kendra Nightingale)
9:30 a.m. Concepts in Microbial Taxonomy and Diversity and Implications for Molecular Detection (Martin Wiedmann)
10:00 a.m. PCR: Fundamentals (Sarita Raengpradub)
10:30 a.m. Break
10:45 a.m. PCR Variations (PCR; Nested PCR; Multiplex PCR; Real-Time PCR; Reverse Transcriptase PCR; Controls and GLPs) (Sarita Raengpradub)
11:15 a.m. Primer Design and Hands-on Primer Design Activity (Kendra Nightingale)
12:00 p.m. Lunch (provided)
1:00 p.m. Introduction to DNA Sequence Databases, Downloading Sequence Data, Alignment Programs (Soraya Chaturongakul and Kendra Nightingale)
1:30 p.m. Design and Order Primers for Custom PCR Assay for Target Organism of Interest (e.g., Salmonella, E. coli O157:H7, L. monocytogenes, Listeria spp. Pseudomonas fluorescence, Paenibacillus spp., Alicyclobacillus)
4:30 p.m. Break for Day
5:00 p.m. Opening Reception (from 5:00 to 6:00 p.m.); Keynote Address TBA; USAFRIMS-Thailand (from 6:00 to 7:00 p.m.); and Dinner (beginning at 7:00 p.m.) Location TBA
Tuesday, March 31st (Lecture Series Day 2; TBA):
8:30 a.m. Fundamentals of DNA Sequencing and 16S rDNA-Based Detection Strategies (Kendra Nightingale)
9:30 a.m. Assay Validation and Approval, Including Use and Misuse of the Sensitivity and Specificity concepts (Mark Carter or Kendra Nightingale)
10:15 a.m. The Steps Before PCR: Sampling and Enrichment, Concentration Procedures, Compositing, True Real Time PCR without Enrichment (Mark Carter or Sarita Raengpradub)
11:00 a.m. Break
11:15 a.m. The Ins and Outs of Selected Commercially Available DNA-Based Detection Assays (Martin Wiedmann)
12:00 p.m. Lunch (provided)
1:00 p.m. Presentations from representatives of companies that manufacture commercial molecular detection kits
2:00 p.m. Question and Answer Session with Company Representatives and Workshop Instructors on Selection of the Molecular Detection Assay that Fits Your Needs
2:30 p.m. Case Studies on Interpretation of Results from Nucleic Acid Based Detection Methods (Including Questions such as “What Does a Positive PCR Assay Mean?” and “How to Deal with a PCR Positive Culture Negative Result?”) / Break-out into Groups and Troubleshoot
3:30 p.m. Complete Post-symposium Evaluations
3:45 p.m. Discussion and Wrap-up Question and Answer Session
4:30 p.m. Break for Day and Dinner on Your Own
Wednesday, April 1st (Hands-on Workshop Day 1; MDL2, second floor of Building K):
8:30 a.m. Prepare Conventional Lysates for PCR Amplification of sigB for Listeria isolates and Lysates for Bax® L. monocytogenes SYBR Green Detection PCR.
Informal Lecture on Cross-contamination of PCR Reactions in During Set-up
10:30 a.m. Set-up sigB PCR Reactions and Begin Thermal Cycling
12:00 p.m. Lunch (provided)
1:00 p.m. Run Gel of sigB PCR Products; Purify sigB PCR Products, Quantify DNA, and Submit for DNA Sequencing
Informal Lecture on Quantification of nucleic acids (Martin Wiedmann)
4:00 p.m. Set-up Bax® L. monocytogenes SYBR Green Detection PCR and Begin Thermal Cycling
4:30 p.m. Break for Day and Dinner on Your Own
Thursday, April 2nd (Hands-on Workshop Day 2; MDL2, second floor of Building K):
8:30 a.m. Interpret and Discus Results from Bax® L. monocytogenes SYBR Green Detection PCR
9:00 a.m. Design Custom PCR Master Mix and Thermal Cycling Conditions
10:30 a.m. Set-up Custom PCR Reactions and Begin Thermal Cycling
12:00 p.m. Lunch (provided)
1:00 p.m. Set-up and Run Gel from Custom PCR
1:30 p.m. Considerations for Application of Molecular Detection Methods (Integration of Molecular and Cultural Methods, PCR Analysis on Enrichments, PCR on Immuno-Magnetic Separation Beads, Quality Control)
2:30 p.m. Interpretation and Troubleshooting of Custom PCR Results
3:30 p.m. Optimization of Custom PCR or Purification of Custom PCR Products and Send for Sequencing (Optional)
4:30 p.m. Break for Day
6:00 p.m. Workshop Closing Dinner at TBA (Shuttle provided); Mixer from 6:00 p.m. to 6:30 p.m. Followed by Dinner at 6:30 p.m.
Friday, April 3rd (Hands-on Workshop Day 3, MDL2, second floor of Building K):
8:30 a.m. TaqMan Real-time PCR to Detect E. coli O157:H7 (ABI, Master Mix Kit)
9:30 a.m. Lecture and Discussion “mRNA Detection” (Sarita Raengpradub and Soraya Chaturongakul)
10:30 a.m. Hands-on Activity to Design TaqMan Primers and Probes (Sarita Raengpradub and Soraya Chaturongakul)
11:15 a.m. Explore Ribosomal Project Database (RDP) through Instructor Assignments (Martin Wiedmann and Kendra Nightingale)
12:00 p.m. Lunch (provided)
1:00 p.m. Assemble and Proofread Sequence Data and Perform BLAST Searches
3:00 p.m. Complete Post-workshop evaluations
3:15 p.m. Optional Wrap-up Discussion Session and Individual Discussion with Instructors
5:00 p.m. Informal Happy Hour, Location TBA
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