Isolation of rare transcripts by representational difference analysis
Author(s) -
Michael O’Neill
Publication year - 1997
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/25.13.2681
Subject(s) - biology , false positive paradox , representational difference analysis , cloning (programming) , computational biology , genome , complementary dna , genetics , isolation (microbiology) , differential display , gene , microbiology and biotechnology , bioinformatics , artificial intelligence , computer science , programming language
Representational difference analysis (RDA) is a powerful technique for cloning the differences between genomes, and has recently been adapted for cloning differentially expressed genes. RDA, like other PCR-based differential screening methods, is prone to the production of false positives. We have identified a major source of false positives in RDA of cDNA and have introduced improvements which minimise their production. These modifications also significantly increase sensitivity, allowing for the isolation of rare differential transcripts from nanogram amounts of mRNA.
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