3D culture hydrogel was transferred to a 1.5 ml centrifuge tube and immersed in RNA lysis buffer, which was placed on dry ice for rapid freezing and then stored at -80 °C. 3D culture hydrogel was homogenized in lysis buffer. Total RNA isolation was performed using a kit following the manufacturer's protocol. Further purification step was performed on the eluted RNA samples. RNA-Seq libraries were prepared following the Smart-seq2 protocol. 1 ng of total RNA was used. cDNA was synthesized and then used in each NexteraXT (Illumina) reaction.
GEO Accession
GSM3040421