title: ShapeTest4_reference for the Bigheart lncRNA
design: We performed SHAPE-Seq for human Bigheart to determine the secondary structure. The RNA transcripts were generated in vitro from a ThermoFisher Gene Synthesis Plasmid with the T7 RNA polymerase. For this the T7 promoter and a sequence specific restriction site was attached to the sequence. For practical reasons, we restricted our approach to RNAs that are no longer than 1000 nucleotides. In vitro SHAPE-Seq was performed according to the protocol by Watters and Lucks15 using the 1M7 reagent, which can achieve a single nucleotide resolution.