Illumina HiSeq 2500 paired end sequencing; Clinical exome sequencing of cells freshly isolated from 12 human colorectal carcinoma patients (tumor endothelial cells, normal colon endothelial cells, PBMCs, n=12 each) in comparison to microdisected tumor DNA (n=11) from corresponding FFPE-tissue blocks
Clinical exome sequencing of cells freshly isolated from 12 human colorectal carcinoma patients (tumor endothelial cells, normal colon endothelial cells, PBMCs, n=12 each) in comparison to microdisected tumor DNA (n=11) from corresponding FFPE-tissue blocks
TECs and NECs were isolated from 12 human CRC patients according to Naschberger et al, JoVE 2018. PBMCs were obtained from whole blood from the same patients and corresponding tumor DNA was extracted from microdissected tumor cells from FFPE-tissue blocks. Total DNA was extracted from TECs or NECs using the QIAamp DNA mini kit (Qiagen). DNA quantity was determined using a Nanodrop 2000c. Extraction of genomic DNA from PBMCs derived from fresh blood was performed by magnetic bead based DNA extraction with the Chemagen chemagic 360 instrument (Perkin-Elmer, Waltham, MA, USA). Tumor DNA was isolated from FFPE tissue after microdissection as previously described (StÃ¶hr et al, Pathobiology 2019). Whole Exome Enrichment and Library Prep was performed with SureSelect version 6 (Agilent).