P-EMBL-79 - P-EMBL-79
About 13000 cells from each fraction were lysed and total RNA isolated using the Acturus PicoPure RNA-Isolation Kit according to the manufacturer's instructions (Arcturus, Mountain View, USA). DNase treatment was performed (Quiagen, Hilden, Germany). RNA quality was controlled with the RNA 6000 Pico LabChip kit (Agilent, Waldbronn, Germany). Total RNA of three individual samples was pooled for each amplification. Linear amplification of RNA was performed by in vitro transcription over two rounds using the Arcturus RiboAmp Kit (Arcturus, Mountain View, USA) according the manufacturer's instructions. Quality and quantity of the amplified RNA was controlled by the RNA nano Lab Chip kit (Agilent) and by the SpectraMAX plus photometer (Molecular Devices, Sunnyvale, USA) at 260 nm. We gained about 80 µg aRNA with a continuous spectrum of RNA length of 200 to 800 base pairs.
Amplification, Extracted product