Exploring Reverse Transcription for DNA MicroArrays
Poster Oct 10, 2005
Miroslava Cuperlovic-Culf, Adrian S. Culf, Dominique Richard, Mark LaFlamme, Daniel Leger and Rodney J. Ouellette
AbstractWe designed 96 un-modified 50-mer DNA oligonucleotides, one for every 2 kb of the longest human genes known: Titin, Nebulin and Obscurin, all expressed in human muscle (Gene length ~100kb). DNA oligonucleotide targets for positive (a-Actin) and negative controls (ß-Actin) for human muscle tissue were included and spotted on epoxide-coated glass slides at 1 - 50µM concentration to create a DNA microarray.
Oligo-dT, random and non-priming strategies were explored for reverse transcription (RT). Results were determined by microarray spot fluorescence analysis.
Our results suggest that random priming is the optimal method for expression analysis of long genes. This method of RT may be appropriate for alternatively spliced genes and for genes without unique probes in the 3’-region.
CiPA Phase 2 Study: validation of an automated microelectrode array (MEA) assay of hiPSC-derived cardiomyocyte electrophysiology for cardiac safety evaluationPoster
These results support the use of hSC-CM and MEA technology for preclinical assessment of proarrhythmic risk within the proposed CiPA paradigm, and, more generally, demonstrate that automation of the CM-MEA assay can achieve high reliability and throughput for cardiac risk assessment in vitro.READ MORE