NextSeq 500 paired end sequencing; Reconstructing the RUNX2 transcriptional hierarchy in thyroid cancer using a multi-layered network- based approach
Reconstructing the RUNX2 transcriptional hierarchy in thyroid cancer using a multi-layered network- based approach
Cells were collected after 10 days from selection. Cells were cultured in DMEM medium supplemented with 10% FBS and 1% P/S. Cells were infected with pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Puro containing either sgRUNX2 or sgNT, then selected with 1ug/ml Puromycin for 3 days. Total RNA was extracted by Maxwell RSC simplyRNA Cells kit (Promega, Madison, Wisconsin, USA) Libraries were obtained starting from 100ng of total RNA following the Illumina Stranded Total RNA preparation Ligation with Ribo-Zero Plus protocol.