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EMBL Master Course

Advanced qPCR Techniques
for Publication Success:

Following MIQE Recommendation

EMBL Heidelberg, Germany Monday 2 July - Friday 6 July 2012
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Preliminary Programme

 
Day 1 - Monday, 2 July 2012
09:00 - 18:00 qPCR Assay Design and Optimization
  An introduction to MIQE
QPCR assay design/Solutions for difficult designs (LNA)
COMPUTER PRACTICAL: QPCR Assay design
Lunch
QPCR assay optimization
LAB PRACTICAL: QPCR Optimisation
Presentation of group challenges
Summary of the day 
 
 
Day 2 - Tuesday, 3 July 2012
09:00 - 18:00 RNA QC Day
  COMPUTER PRACTICAL: Analyse optimization data
The effect of RNA degradation on RT-qPCR
Lunch
The effect of inhibitors on RT-qPCR
LAB PRACTICAL: Standard RNA quality control assessments
LAB PRACTICAL: Checking RNA integrity (5’/3’) and quantification
LAB PRACTICAL: Assessing sample quality (SPUD)
Analysis of QC data
Summary of the day
 
 
Day 3 - Wednesday, 4 July 2012
09:00 - 18:00 RT and Normalisation Day
  RT- A Necessary Evil
PRACTICAL: RT reactions – priming comparison
Lunch
Ears, BEERs and Normalisation
Reference gene selection and normalisation
COMPUTER PRACTICAL: Selecting reference genes/system
Comparison of normalization approaches for microRNA normalization
Summary of the day
 
 
Day 4 - Thursday, 5 July 2012
09:00 - 18:00 Gene Expression Analysis Day
  Genomic Regulation of Gene Expression
Quantification strategies and efficiency determination
PRACTICAL: qPCR gene expression experiments
Lunch
Data analysis using REST Lessons from the MMR scandal
QC before publication
Data analysis – RT comparison
Summary of the day
 
 
Day 5 - Friday, 6 July 2012
09:00 - 16:00 Data Analysis and Troubleshooting Day
  Data analysis – some thoughts on biostatistics
Lunch
COMPUTER PRACTICAL: Data analysis iron model using REST/ geNorm/ qBASE/ GenEx
COMPUTER PRACTICAL:Interpretation of ZFN data
Troubleshooting, summary of the course
End of course