NucleoSpin RNA, Mini kit for RNA purification (According to manufacturer protocol), (Machery-Nagel) Performed by Single Cell Discoveries (Netherlands) 1. Pre-quality checks to measure RNA concentration and RNA quality 2. Performing RT to generate cDNA 3. Pooling of all samples (barcodes are now integrated within the cDNA molecules) 4. Amplification of the material through an IVT reaction (linear amplification), which generates amplified RNA 5. QC: bioanalyzer of the amplified RNA 6. Library preparation to generate a sequenceable cDNA library (aRNA) 7. QC: bioanalyzer of the final cDNA library