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Updated: Jun 3, 2026

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
Published on: June 14, 2021
Generation of a polymerase chain reaction renewable source of subtractive cDNA
1NCI-Navy Medical Oncology, Bethesda, MD.
Abstract:
Differential (+/-) first-strand cDNA screening methods identify clones corresponding to mRNAs that are expressed at a higher level in one of a pair of phenotypically different cells. This approach is limited by the fact that screening of libraries with labeled first-strand cDNAs synthesized from unfractionated mRNA can detect clones containing sequences representing approx 0.1% or more of the complexity of mRNA (i.e., mRNAs present at greater than about 200 copies per cell since a typical mammalian cell line contains approx 250,000 mRNAs).
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