Illumina HiSeq 2000 paired end sequencing; Analysis of human enteroids and colonoids (3D organoid models from small intestine and colon) exposed to chemotherapy agent 5-fluorouracil (5FU) in a dose-repeated exposure across three doses (10,100 and 1000uM) and sampled at three points (24, 48 and 72h) since initial exposure. Organoids were obtained from healthy human tissues.
Analysis of human enteroids and colonoids (3D organoid models from small intestine and colon) exposed to chemotherapy agent 5-fluorouracil (5FU) in a dose-repeated exposure across three doses (10,100 and 1000uM) and sampled at three points (24, 48 and 72h) since initial exposure. Organoids were obtained from healthy human tissues.
Remove medium from each well and add 200 µL of QIazol Lysis reagent (Qiagen, The Netherlands) into the wells to dissociate the matrigel matrix and transfer each sample to the respective tubes. Repeat this step 2 or 3 times until all organoids are collected to a total volume of QIazol Lysis reagent of 700 µL. Performed using miRNeasy Mini Kit (Qiagen, The Netherlands), following manufacturer\u2019s protocol for Animal Cells including a DNase treatment. Samples with purified RNA were prepared for sequencing using Lexogen SENSE mRNA- seq protocol and library preparation kit (Lexogen, The Netherlands).