title: Detection of antisense transcripts in transgenic plants by RT-PCR.
abstract: 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.
authors.affiliation: Cell and Molecular Genetics Department, Scottish Crop Research Institute, Invergowrie, Dundee, UK.
authors.forename: J W
authors.initials: JW
authors.lastname: Brown
authors.forename: G G
authors.initials: GG
authors.lastname: Simpson
authors.forename: G
authors.initials: G
authors.lastname: Clark
authors.forename: J
authors.initials: J
authors.lastname: Lyon
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.forename: C
authors.initials: C
authors.lastname: Simpson
mesh_terms: D001483:Base Sequence; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016372:RNA, Antisense / Q000302:isolation & purification*; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D016372:RNA, Antisense; D017413:Ribonucleoprotein, U2 Small Nuclear