ArticleBMC genomics2005
A robust method for the amplification of RNA in the sense orientation.
Article in BMC genomics, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- LINCATRA: Two-cycle method to amplify RNA for transcriptome analysis from formalin-fixed paraffin-embedded tissue.Heliyon · 2024Article
- The successes and future prospects of the linear antisense RNA amplification methodology.Nature protocols · 2018Article
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- Selective amplification of Brucella melitensis mRNA from a mixed host-pathogen total RNA.BMC research notes · 2010Article
- Model-based probe set optimization for high-performance microarrays.Nucleic acids research · 2009Article
- Optimization of experimental design parameters for high-throughput chromatin immunoprecipitation studies.Nucleic acids research · 2008Article
- Insulin-like growth factor binding protein 7 mediates glioma cell growth and migration.Neoplasia (New York, N.Y.) · 2008Article
- Terminal Continuation (TC) RNA amplification enables expression profiling using minute RNA input obtained from mouse brain.International journal of molecular sciences · 2008Article
- Multiplex preamplification of specific cDNA targets prior to gene expression analysis by TaqMan Arrays.BMC research notes · 2008Article
- FK506 binding protein mediates glioma cell growth and sensitivity to rapamycin treatment by regulating NF-kappaB signaling pathway.Neoplasia (New York, N.Y.) · 2008Article
- Article
- Evaluation of global RNA amplification and its use for high-throughput transcript analysis of laser-microdissected endosperm.International journal of plant genomics · 2007Article
- Options available for profiling small samples: a review of sample amplification technology when combined with microarray profiling.Nucleic acids research · 2006Review
- Advances in molecular phytodiagnostics - new solutions for old problems.European journal of plant pathology · 2006Review
- Limitations of mRNA amplification from small-size cell samples.BMC genomics · 2005Article
- RNA amplification for successful gene profiling analysis.Journal of translational medicine · 2005Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSmall quantities of RNA (1-4 microg total RNA) available from biological samples frequently require a single round of amplification prior to analysis, but current amplification strategies have limitations that may restrict their usefulness in downstream genomic applications. The Eberwine amplification method has been extensively validated but is limited by its ability to produce only antisense RNA. Alternatives lack extensive validation and are often confounded by problems with bias or yield attributable to their greater biological and technical complexity.
resultsTo overcome these limitations, we have developed a straightforward and robust protocol for amplification of RNA in the sense orientation. This protocol is based upon Eberwine's method but incorporates elements of more recent amplification techniques while avoiding their complexities. Our technique yields greater than 100-fold amplification, generates long transcript, and produces mRNA that is well suited for use with microarray applications. Microarrays performed with RNA amplified using this protocol demonstrate minimal amplification bias and high reproducibility.
conclusionThe protocol we describe here is readily adaptable for the production of sense or antisense, labeled or unlabeled RNA from intact or partially-degraded prokaryotic or eukaryotic total RNA. The method outperforms several commercial RNA amplification kits and can be used in conjunction with a variety of microarray platforms, such as cDNA arrays, oligonucleotide arrays, and Affymetrix GeneChip arrays.
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