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.
The immune system is a striking example of an integrated information system, engaged in coordinated host-protective activities. Organs-on-chip approach (OOC) models allow the direct simultaneous observation of hundreds of different cells, moving, interacting and responding to signals coming from the microenvironment nearby, that give access to a number of parameters describing the system that must be properly measured and elaborated.READ MORE