Amplification of RNA transcripts using terminal continuation.
作者： Shaoli Che , Stephen D Ginsberg
DOI: 10.1038/LABINVEST.3700005
关键词: RNA spike-in 、 RNA 、 Molecular biology 、 Rapid amplification of cDNA ends 、 RNA silencing 、 Intron 、 Antisense RNA 、 RNA-dependent RNA polymerase 、 Non-coding RNA 、 Biology
摘要: A new methodology has been developed to amplify RNA from minute amounts of starting material. Specifically, an efficient means second-strand (ss) cDNA synthesis using a sequence-specific 'terminal continuation' (TC) method is demonstrated. An promoter attached the 3' and/or 5' region utilizing TC mechanism. The orientation amplified RNAs 'antisense' or novel 'sense' orientation. amplification utilized for many downstream applications including gene expression profiling, microarray analysis, and library/subtraction library construction. Synthesized sense TC-amplified can also be used as template in vitro protein translations proteomic applications. offers high sensitivity, flexibility, throughput capabilities. likely mechanism that primer binds preferentially GC-rich CpG islands flanking regions DNA contain sequences. Following amplification, large proportion genes assessed quantitatively evidenced by bioanalysis analysis mouse human postmortem brain tissues.
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