GSM8230065: LAM-27/RBJ; Homo sapiens; RNA-Seq
Tissue sections were mounted on PEN Membrane Glass Slides, deparaffinised and stained using Cresyl Violet. LCM was carried out using a Zeiss AxioImager Z1. Microdissected samples were captured by AdhesiveCap 500ul microcentrifuge tubes. RNA extraction was performed by using RNeasy DSP FFPE Kit (QIAGEN, ID: 73604). Genomic DNA was eliminated using, RQ1 RNase-Free DNase (Promega, M6101) and clean-up performed using Monarch® RNA Cleanup Kit (Biolabs, New England, T2030L). . The library preparation and RNA sequencing was performed by Birmingham Genomic centre using Lexogen QuantSeq 3' mRNA-Seq Library Prep Kit FWD from Illumina. Libraries were quantified using a PicoGreen Quant-iT kit and sized with a D1000 tape For sequencing, libraries were pooled in equal volumes according to their molarity. These were then quality checked using the Agilent TapeStation DNA 1000 tape and DNA HS Qubit. Pooled libraries was taken into sequencing at 2nM and loaded on the NovaSeq 6000 at 400pM with an SP 100 cycle flowcell.