Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: 50 µl extraction buffer (PicoPure RNA Isolation Kit (Applied, Biosystems, Carlsbad, USA)) was added to the captured LCM material followed by incubation for 45 min at 60 °C to allow de-crosslinking. RNA was extracted with the PicoPure RNA Isolation Kit. 8 µL of the RNA solution was denatured, treated with DNase and reverse transcribed into cDNA using a combination of random and Oligo(dT) primers. The cDNA was amplified with the Qiagen Repli-G WTA (whole transcriptome amplification) Single Cell Kit (Qiagen, Hilden, Germany) using a mix of random and poly-A primers. unstranded RNA-seq