Bx[RP] - similar to Ubx[RP] and kuz[RP], except for the following differences: homozygous mutant adult flies were homogenized and RNA was extracted using Trizol. 5 μg of RNA was DNase treated and reverse transcribed using random hexamers. rt-PCRs were done using 35 cycles for mRNA and intermediates.
Cell culture: RNA was collected from transfected cells using Trizol. 5 μg of RNA was treated with Turbo DNase [Ambion] for 45 min before cDNA synthesis using SuperScript III [Life Technology] with random hexamers. rt-PCRs were done using AccuPrime™ Pfx DNA polymerase [ThermoFisher Scientific] with standard protocol using 26 cycles and primers that were specific to minigene construct. All primers with descriptions can be found in