Abstract
The debut of RNA-polymerase cycling reaction (RNA-PCR) has promised to provide linear amplification of a reproducible mRNA library from as few as 20 single cells (1). By incorporating a RNA promoter element during the synthesis of double-stranded complementary DNA (cDNA) templates, a poly(A+) RNA library can be generated and reamplified from the templates in the same conformation and composition as its mRNA origins (Fig. 1). Using microarray analysis, the RNA-PCR-derived poly(A+) library has been proven to contain above 97% of the original poly(A+) RNA population and maintain 88±4% linear correlationship to the populationary ratio of each RNA species (Chapter 12). It has also been tested to generate a full-length mRNA library from as few as 20 homologous tissue cells (2-pg mRNAs) for profiling cancer stages in vivo.
An illustration of using RNA-PCR-derived poly(A+) RNA for Northern blot analysis. The mRNA generated in step D can be fractionated on a formaldehydeagarose gel for specific gene detection.
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© 2003 Humana Press Inc., Totowa, NJ
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Lin, SL. (2003). Single-Cell mRNA Library Analysis by Northern Blot Hybridization. In: Ying, SY. (eds) Generation of cDNA Libraries. Methods in Molecular Biology™, vol 221. Humana Press. https://doi.org/10.1385/1-59259-359-3:169
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DOI: https://doi.org/10.1385/1-59259-359-3:169
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