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Promises of studying the transcriptome
• RNA is easy to replicate and synthetise
• RNA-detection can be more versatile than protein-directed diagnostics
• Capture an additional level of complexity above the simple genome sequence
Promises of transcriptomics
• Analysis of gene expression in its entirety allows detection of broad coordinated trends which cannot be discerned by more targeted assays.
• Map the unique gene expression patterns that define cell types and diseases
• Generate a comprehensive picture of what genes are active in specific pathogenetic states
Challenges
• Pre-analytical variation
o RNAdegradation
o Sample contamination (RT or PCR inhibitors)
• Analytical variation
o Accuracy – Reproducibility – Sensitivity – Specificity of assays o Absolute quantification versus relative expression levels
o Datanormalisation
• Biological variation
o RNAbiology:dynamicandcomplex
o Whatisagoodcontrolgroupinendometriosis?
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