Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization

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ID: 301795
1993
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Abstract
Differential display has been developed as a tool to detect and characterize altered gene expression in eukaryotic cells. The basic principle Is to systematically amplify messenger RNA5 and then distribute theIr 3′ termini on a denaturing polyacrylamide gel. Here we provide methodological details and examine in depth the specificity, sensitivity and reproducibIlity of the method. We show that the number of anchored 01190- dT primers can be reduced from twelve to four that are degenerate at the penultimate base from the 3′ end. We also demonstrate that using optimized conditions described here, multiple RNA samples from related cells can be displayed simultaneously. Therefore process-specific rather than cell-specific genes could be more accurately identified. These results enable further streamlining of the technique and make It readily applicable to a broad spectrum of biological systems.
Reference Key
openalex_W1995522877 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Liang Peng, Arthur B. Pardee
Journal Nucleic Acids Research
Year 1993
DOI
10.1093/nar/21.14.3269
URL
https://doi.org/10.1093/nar/21.14.3269
Keywords Keywords not found

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