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The validation study design consisted of an assessment of the microarray chip performance using the MessageAmp and FairPlay labelling kits.
After extraction, RNA was amplified by using the MessageAmp II aRNA Amplification Kit (Ambion).
500 ng total RNA was amplified using the MessageAmp modified Eberwine linear amplification procedure (Applied Biosystems).
Both sample and reference RNAs were amplified using the MessageAmp II aRNA amplification kit (Ambion, Austin, TX).
Synthesis and linear amplification of cRNA were accomplished by transcription in vitro using the MessageAmp RNA kit (Ambion).
Total RNA (2 3 ng) was amplified using the MessageAmp II aRNA Amplification Kit (Ambion, USA) according to manufacturer's recommendations.
mRNA was linearly amplified using the MessageAmp II-Bacteria kit (Ambion) according to the manufacturer's instructions.
RT, second strand synthesis, cDNA purification and in vitro transcription with Biotin-11-UTP were performed using the MessageAmp II aRNA Amplification Kit (Ambion).
Only high quality RNA was subjected to two rounds of linear amplification using the MessageAmp II aRNA Amplification Kit (Ambion, USA) according to the manufacturer's instructions and amplified RNA (aRNA) was quantified using the RNA 6000 Pico Assay Kit.
Biotin-labeled cDNA was generated using the MessageAmp II kit (Ambion, Austin, TX).
Total RNA isolated from these cells was amplified using the MessageAmp Premier protocol.
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