A robust method for the amplification of RNA in the sense orientation.

摘要: Small quantities of RNA (1–4 μg total RNA) available from biological samples frequently require a single round amplification prior to analysis, but current strategies have limitations that may restrict their usefulness in downstream genomic applications. The Eberwine method has been extensively validated is limited by its ability produce only antisense RNA. Alternatives lack extensive validation and are often confounded problems with bias or yield attributable greater technical complexity. To overcome these limitations, we developed straightforward robust protocol for the sense orientation. This based upon Eberwine's incorporates elements more recent techniques while avoiding complexities. Our technique yields than 100-fold amplification, generates long transcript, produces mRNA well suited use microarray Microarrays performed amplified using this demonstrate minimal high reproducibility. describe here readily adaptable production antisense, labeled unlabeled intact partially-degraded prokaryotic eukaryotic outperforms several commercial kits can be used conjunction variety platforms, such as cDNA arrays, oligonucleotide Affymetrix GeneChip™ arrays.