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Detection of antisense transcripts in transgenic plants by RT—PCR
Author(s) -
Brown John W.S.,
Simpson Gordon G.,
Clark Gillian,
Lyon Jackie,
Kumar Amar,
Simpson Craig
Publication year - 1993
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1993.04050883.x
Subject(s) - reverse transcriptase , primer (cosmetics) , oligonucleotide , sense (electronics) , microbiology and biotechnology , biology , rna , antisense rna , gene , polymerase chain reaction , transgene , reverse transcription polymerase chain reaction , gene expression , primer dimer , genetics , chemistry , multiplex polymerase chain reaction , organic chemistry
A reverse transcriptase—polymerase chain reaction (RT—PCR) where one oligonucleotide primer is end‐labelled has been used to analyse expression in transgenic plants carrying antisense gene constructs. Specific detection of both sense and antisense RNA transcripts of the spliceosomal protein gene, U2B″, was achieved using the same pair of oligonucleotide primers. To maintain specificity, a reaction step in which reverse transcriptase was inactivated and RNA digested was found to be essential.