Fig. 1

T7SL IVT method. a Comparison between T7oligo(dT) (left) and T7SL IVT (right) methods. In the Eberwine method, the reverse transcription reaction is performed using a T7oligo(dT) primer, resulting in a first-strand cDNA containing the T7 promoter. The T7 RNA polymerase is added for in vitro transcription of the purified cDNA and antisense aRNA is obtained. For the second method (T7SL IVT), the reverse transcription reaction is performed using random primers and the second-strand cDNA is obtained by a DNA polymerase reaction with the T7SL primer, which also contains the T7 promoter. After cDNA purification, in vitro transcription with the T7 RNA polymerase is performed, producing sense aRNA. b Distribution of RNA lengths from IVT samples and poly(A) + RNA (control), quantified by the BioAnalyzer 2100 equipment (Agilent). c Median length of poly(A) + RNA and aRNAs from the IVT samples. d aRNA yield obtained from the two IVT methods