Pre-Conference Tutorial* 

miRNA Analysis: From Sample to Discovery

Thursday, March 29 

8:00-9:00 Registration for Pre-Conference Tutorial* 

9:00-12:00 Pre-Conference Tutorial 
miRNA Analysis: From Sample to Discovery 
Instructor: Tim Davison, Ph.D., Senior Scientist, Services Division, Asuragen Inc. 
The discovery of micro-RNAs (miRNAs) and the growing appreciation of the importance of miRNAs in the regulation of gene expression are driving increasing interest in miRNA expression profiling. Early studies have suggested prominent roles for these genetically encoded regulatory molecules in a variety of normal biological processes and diseases, particularly cancer. However, the field of miRNA expression profiling is in its infancy. Several factors, including the small size, the unknown but limited number of miRNAs, and the tissue-to-tissue and tissue-to-disease state variability in miRNA expression, make the adaptation of microarray technology to the evaluation of miRNA expression nontrivial. In this tutorial we discuss the unique features of miRNA experiments and analysis from sample preparation, choice of expression platform and subsequent analysis. Depending on time and interest of the workshop participants, we will discuss topics including miRNA prediction, miRNA target prediction and the miRNA therapeutic/diagnostic discovery pipeline.

• miRNA sample preparation: 
  • miRNA isolation – various RNA isolation technologies are discussed in the context of recovery of the small RNA fraction. 
  • Enriching miRNA content – this is an optional procedure depending on the platform for analysis. The benefits and pitfalls of enrichment are discussed. 
  • How much miRNA do I have in my sample? – Many downstream platform technologies and analyses have variable performance depending on the absolute and relative quantities of miRNA in your sample. The topic is introduced for purposes of awareness and discussion. 

   

• miRNA analysis platforms: 
  • miRNA qRT-PCR expression analysis 
    • How quantitative is my assay? 
    • Can I compare CT within sample vs. between samples? 
    • Are there any “housekeeping” miRNA to use as a reference standard? 
  • miRNA Array expression analysis 
    • Review of available array technologies for miRNA 
    • 1-color vs. 2-color 
    • How do I normalize my data? Which statistical analysis algorithms & tools from mRNA array analysis can I port over to miRNA array analysis? 
  • miRNA Platform concordance 
    • How should I assess concordance between array and PCR technologies? 
    • Specificity and Sensitivity comparison for PCR and array technologies 

   

• miRNA annotation: 
  • Summary of present algorithms for miRNA prediction 
  • How does a miRNA sequence become a member of Sanger? 

   

• miRNA target prediction: 
  • Summary of present algorithms for target prediction 
  • What are the present options? 
  • Do they agree? If not why not and what should I do? 

   

• miRNA therapeutic/diagnostic discovery pipeline: 
  • Review of published potential miRNA therapeutic targets 
  • What are the practical limitations to miRNA as biomarkers? 
  • What are the benefits of miRNA as biomarkers?