DIRECT ISOLATION OF TRANSPOSON INSERTIONS TAGGING TRANSCRIBED PORTIONS OF GENES

摘要

The invention concerns a method for the identification of transposon insertions within a transcribed portion of a gene of interest in an organism or tissue of interest, said method comprising the following steps: isolating RNA from the organism or tissue of interest; hybridizing the isolated RNA to one or more polymer molecule(s) containing nucleobases capable of hybridization with at least a part of the transposon sequence in a chimeric RNA gene transcript; performing a reverse transcription reaction primed by said polymer molecules; optionally amplifying the cDNA resulting from the reverse transcription reaction; analysing the obtained cDNA fragments by one or more of the following methods: d) analysing the obtained cDNA fragments for segregation of fragments; e) using the fragments as probes or targets in nucleic acid hybridization assays; f) sequencing of the obtained cDNA fragments; various applications of said method as well as kits.