High-accuracy amplification of nanogram total RNA amounts for gene profiling.

摘要: Microarray-based gene profiling of laser-assisted microdissected tissues or clinical biopsies is still a challenge since the amount total RNA in such samples limited and amplification mandatory. Representative mRNA highly dependent on reverse transcription reaction, which error prone, number cycles. To improve accuracy amplification, we optimized, combined, tested different strategies for Affymetrix oligonucleotide array hybridization. We demonstrate that protocols differ significantly quality amplification. reproducibility our optimized protocol setting, analyzed RNAs from laser-assisted, cells human prostate tissues. On basis these results, recommend standard reaction small-sample-transcriptome experiments as part Minimal Information about Microarray Experiment (MIAME) set standards.