Following MIQE Recommendations

Anders Bergkvist/Tania Nolan

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

18.00 Dinner at EMBL
19.00 Get Together
20.30 Bus Shuttle to isg Hotel

Tania Nolan/Jim Huggett/Michael Pfaffl 

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

18.00 Dinner at EMBL
19.00 Bus Shuttle to isg Hotel

Jo Vandesompele/Tania Nolan

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

18:00 Bus to isg, free evening

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

18:00 Bus to isg hotel
18.45 Bus isg Hotel -> Downtown
19:00 Dinner downtown

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

16:00 End of the course