Wednesday, 30 April 2014
|10:45||Peripheral Whole Blood Transcriptomes of Asthmatic Individuals Undergoing Allergen Inhalation Challenge
Scott Tebbutt, Associate Professor, University of British ColumbiaAllergen inhalation challenge can be a suitable model to explore the dynamics of immune cells and to elucidate the mechanism of allergic asthma in humans. We performed RNA-seq transcriptomic analysis on Illumina rRNA/globin-depleted stranded cDNA libraries generated from blood leukocyte samples donated by asthmatic individuals undergoing allergen inhalation challenge.
|11:30||Molecular Indexing For Improved RNA-Seq Quantitation and Analysis
Leah Matzat, Senior Scientist, Bioo ScientificMost modern methods for NGS library prep require the use of enzyme processing, such as DNA polymerase reactions, which can introduce errors in the form of incorrect sequence and misrepresented copy number. Conventional RNA sequencing library construction involves the ligation of a population of cDNA molecules with adapters prior to amplification and sequencing. With Molecular Indexed™ libraries, each molecule is tagged with a molecular index randomly chosen from ~10,000 combinations so that any two identical molecules become distinguishable (with odds of 10,000/1), and can be independently evaluated in later data analysis. Analysis using molecular indexing information provides an absolute, digital measurement of gene expression levels, irrespective of common amplification distortions observed in many RNA-Seq experiments.