Illumina NovaSeq 6000 paired end sequencing; Analysis of human healthy enteroids and colonoids (3D organoid models from small intestine and colon) exposed to chemotherapy agent Doxorubicin (DOX) sampled at four doses (1,10, 30 and 60uM) and three time points (24, 48 and 72h).
Analysis of human healthy enteroids and colonoids (3D organoid models from small intestine and colon) exposed to chemotherapy agent Doxorubicin (DOX) sampled at four doses (1,10, 30 and 60uM) and three time points (24, 48 and 72h).
Remove medium from each well and add 200 L 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 L. Performed using miRNeasy Mini Kit (Qiagen, The Netherlands), following manufacturer's 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).