Gene Expression Profiling Using the Terminal Continuation RNA Amplification Method for Small Input Samples in Neuroscience

The process of RNA amplification is a stepwise series of molecular manipulations intended to amplify transcriptomic signals from small quantities of starting materials, including single cells and homogeneous populations of individual cell types for microarray analysis and other high-throughput downstream genetic approaches. An RNA amplification methodology named terminal continuation (TC) RNA amplification has been developed by our group to amplify RNA from small starting material inputs. In brief, an RNA synthesis promoter is attached to the 3′ and/or 5′ region of the isolated population of cDNAs utilizing the TC approach. Amplified RNAs are in either an antisense or a sense orientation depending on the placement of the T7 RNA polymerase promoter sequence. TC RNA amplification is utilized for many downstream applications including gene expression profiling and cDNA library/subtraction library construction, among others. Input sources of RNA can originate from a myriad of in vivo and in vitro tissue sources. Notably, fresh, frozen, and fixed tissues can be employed for TC RNA amplification, enabling precise and reproducible cell type and tissue specificity of downstream transcriptome-based assessments.