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POSTER
Detection of Prostate Cancer BioMarkers by Expression Microarray Analysis of
TransPlex® WTA2 Amplified FFPE Tissue RNA
Ken Heuerman and Brian Ward
Sigma Life Science
Sponsored by Sigma-Aldrich
Published in
September 2010
2010
(p.6)
DOI: 10.2144/000113526
Sponsored,vendor-submitted protocol
Sigma Life Science
Sponsored by Sigma-Aldrich
Published in
September 2010
2010
(p.6)
DOI: 10.2144/000113526
Sponsored,vendor-submitted protocol
The Transplex® Complete Whole Transcriptome Amplification (WTA2) Kit has been
enhanced to effectively amplify damaged RNA and low input amounts of
RNA.Total RNA samples isolated from matched
FFPE(formalin-fixedparaffin-embedded) and frozen prostate tissues (malignant
and normal) were amplified with the Transplex® Complete WTA2 Kit and
analyzed on Agilent Whole Genome expression microarrays. A call rate of >65%
was observed for combined FFPE and frozen expression differentials
(Malignant/Normal), with >90% commonality between the FFPE and frozen
datasets. Amplification reproducibility of experimental replicates was
demonstrated by principal component analysis.Pair-wise comparison of FFPE
malignant and normal expression(p<0.0001) resulted in detection of 66 known
prostate cancer biomarkers. Similar microarray pair-wise analysis of the
original matched FFPE RNAs amplified with the Nugen WT-Ovation™FFPE Kit at
significantly lower level of confidence(p<0.05) revealed only a single array
feature, disallowing a comprehensive comparison with Transplex WTA2.These
results confirm the utility of the Transplex WTA2Kit for amplification of
damaged DNA.
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