Transcriptional Profiling of Small Samples in the Central Nervous System
RNA amplification is a series of molecular manipulations designed to amplify genetic signals from small quantities of starting materials (including single cells and homogeneous populations of individual cell types) for microarray analysis and other downstream genetic methodologies. A novel methodology named terminal continuation (TC) RNA amplification has been developed in this laboratory to amplify RNA from minute amounts of starting material. Briefly, an RNA synthesis promoter is attached to the 3′ and/or 5′ region of cDNA utilizing the TC mechanism. The orientation of amplified RNAs is “antisense” or a novel “sense” orientation. TC RNA amplification is utilized for many downstream applications, including gene expression profiling, microarray analysis, and cDNA library/subtraction library construction. Input sources of RNA can originate from a myriad of in vivo and in vitro tissue sources. Moreover, a variety of fixations can be employed, and tissues can be processed for histochemistry or immunocytochemistry prior to microdissection for TC RNA amplification, allowing for tremendous cell type and tissue specificity of downstream genetic applications.
Keywordsexpression profiling functional genomics IVT (in vitro transcription) microarray postmortem human brain RNA amplification
I thank Shaoli Che, MD, PhD; Melissa Alldred, PhD; Irina Elarova; Shaona Fang; and Krisztina M. Kovacs for expert technical assistance Support for this project comes from the NINDS (NS43939, NS48447) and NIA (AG10668, AG14449, AG17617, AG09466) and Alzheimer's Association.
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