qPCR Application Workshop
by TATAA Biocenter
logo TATAA
Two types of workshop will be perfomed at the qPCR 2005 Event:
One will be like a classical qPCR Application Workshop organized by the TATAA Biocenter in Sweden (www.tataa.com), and the other is a new type of qPCR Matrix Workshop where people rotate and can visit 6 different workshop sessions. Both qPCR Workshop typess will be held from afternoon 7th till late afternoon 9th September 2005 and will be divided in two different workshop types.
The students can visit either the   TATAA Workshop   or   the qPCR Matrix Workshop.

=> TATAA workshop is fully booked by 20 people
=> please participate in the
qPCR Matrix Workshop

qPCR Application Workshop by TATAA Biocenter
Freising 7-9th September 2005

Practical room number 2 - Plan of room P2

This workshop is aimed at giving participants a deep and objective understanding of real-time quantitative PCR and its applications. The courses are intended for persons considering working with qPCR or scientists currently working with qPCR seeking a deeper understanding.

The course covers all aspects in qPCR, from sample preparation to data analysis and is held during 3 days. The course is approximately 50% hands-on as is limited to 20 participants, resulting in very interactive teaching and everybody given the opportunity to try the instrumentation.

Examples of topics covered in the workshop:

·    Basic Principles of PCR and qPCR
·    Comparison of different detection technologies
·    Applications of qPCR
·    Probe and Primer design
·    Data Analysis
·    Relative Quantification-considerations and limitations
·    Experimental Design
·    Reverse Transcription
·    Extraction methods
·    Multiplex considerations

After the course participants will be able to plan and perform qPCR experiments themselves, as well as interpret and analyze data.
Preliminary Schedule for TATAA Biocenter qPCR workshop
Freising 7-9th September 2005

Practical room number 2 - Plan of room P2

The course is focused on practical issues for qPCR and are partly hands-on, performed by the course participants in the lab (marked in blue). Preliminary agenda for download   TATAA-qPCR-WS.pdf
Lunch, coffee and snacks are included in the course fee.

Wed. 7th
Day 1  -  Basic qPCR
Basic PCR and qPCR theory and applications
  • Amplification and detection
  • Detection chemistries
  • Selected applications
qPCR experiment by participants
  • Display of various instrument platforms
  • Demonstration of qPCR software
  • Practical considerations when preparing PCR reactions
  • Programming qPCR machines
Coffee Break
Primer and probe design and considerations
  • What does primer design affect?
  • What are primer dimers?
  • How do we minimize formation of primer dimers?
  • Design of Molecular Beacons and TaqMan probes
Data analysis
  • How does qPCR software process the data?
  • How are standard curves used and created?
  • How are melt curves used?
  • Principle of quantification using standard curves
  • Principle of relative quantification
Analysis of performed qPCR experiments
Discussion and Q&A
End of qPCR workshop day 1

Thu. 8th
Day 2  -  Advanced qPCR.  Quantification, Normalization and experimental design
qPCR quantification strategies
  • standard curves
  • relative quantification
  • how to compensate for inhibition in biological samples
Normalization of qPCR data
  • What levels of normalization can be used?
  • How to choose a good reference gene?
Coffee Break
Experiment comparing different quantification strategies
  • relative and standard curve quantification
  • different efficiency calculations/assumptions
Optimization of qPCR protocols
  • What parameters can/should be optimized?
  • An optimization strategy
Quantification calculation examples
  • what effect will efficiency have on quantification
  • quantification methods, and equations
Coffee Break
Analysis of experimental data
  • differences in quantifications strategies
  • effect of efficiency estimations on results
  • calculations of relative abundance of genes
  • pros and cons of different methods
Discussion and Q&A
End of qPCR workshop day 2

Fri. 9th
Day 3  -  Advanced qPCR:  Sample Preparation and reverse transcription
Principle of RT and different RT priming strategies
  • Pros and cons of different methods
Principle of RNA and DNA extraction
  • How it works
  • Available methods and products suitable for qPCR
  • Practical considerations
Coffee Break
Reverse transcription experiment using different priming methods
  • Oligo(dt)
  • Random Hexamers
  • Gene specific primers
qPCR experiment evaluating RT using the generated cDNA
  • Is there a best RT priming method?
Quality Control in qPCR using Kinetic Outlier Detection
  • How to detect samples with significant inhibition
Coffee Break
SNP detection. Multiplexing possibilities and problems
  • qPCR for SNP/mutation detection. What alternatives are there?
  • Multiplex optimization
Analysis of experimental data
  • Which priming method for RT is best?
  • How should experiments be planned to take RT priming into consideration?
Probes and Dyes
  • What dyes/quenchers are typically used in qPCR
  • How to measure the maximum fluorescence available in a dual-labelled probe
Discussion and Q&A
End of qPCR workshop day 3

any questions =>  please contact the organizer Neven Zoric via  Neven.Zoric@tataa.com