Sequence-independent Selective Amplification of mRNAs over rRNAs
Poster Nov 29, 2010
John Arrand1, Sim Sihota1, Wenbin Wei1 and Guido Krupp2
Standard mRNA amplifications for "All-Exon" microarrays and for bacterial RNAs are impossible with small samples (< 100 ng total RNA) and with
degraded RNAs, because removal of rRNAs must precede universal, non-selective RNA amplification (Affymetrix and Ambion kits, respectively).
This pre-treatment with magnetic beads is cumbersome, requires high amounts of starting material (>0.5 µg total RNA), is not universal for all species
and degraded RNAs are not suitable. Omission of this step results in lost sensitivity .
However, AmpTec's unique TRinucleotide primer strategy renders reverse transcriptions selective for mRNA (fragments) and it provides sequenceindependent, preferential 3'-proximal priming. This eliminates the rRNA removal step, thus high mRNA amplifications (input of <1 ng total RNA
yields >10 µg amplified RNAs), and the use of degraded RNAs are possible. A first Exon Array study with saliva RNA (<4ng of degraded RNA) is now in press .
 Pepper et al., 2007 “A Core Lab Case Study: Exon Array Challenges and Opportunities” Affymetrix Application Note.
 Hu et al., 2008 (Wong lab at UCLA) “Exon-level expression profiling: a comprehensive transcriptome analysis for oral fluids.”
Clinical Chemistry 54:5 (E-publication ahead of print).
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
Inhibition of The Auto-inflammation Suppressor Protein ISG15 Triggers Preeclampsia by Blocking Trophoblast Migration and InvasionPoster
In summary, ISG15 expression levels are crucial for trophoblast morphology and function (migration/invasion). By blocking trophoblast invasion, reduced ISG15 levels could contribute to impaired spiral artery transformation that reduces utero-placental blood flow in preeclampsia. Thus, agents inducing ISG15 expression are likely to be therapeutic in preeclampsia.
Immunotherapy on Chip: “Under the Microscope” Drug-induced Modulation of the Immune ResponsePoster
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