NextSeq 500 paired end sequencing; Transcriptional dysregulation associated with chromatin accessibility of iPSC and hepatocytes derived from a Cornelia de Lange syndrome patient (sWGS)
Transcriptional dysregulation associated with chromatin accessibility of iPSC and hepatocytes derived from a Cornelia de Lange syndrome patient (sWGS)
Peripheral Blood Mononuclear cells (PBMCs), freshly prepared from the patient blood sample, were reprogrammed using CytoTune-iPS2.1 Sendai Reprogramming Kit (Invitrogen).  Briefly, 5x105 PBMCs were used for reprogramming and more than 20 iPSC-like cell colonies were grown after 3 weeks of culture.  Out of these cell colonies,  several symmetric and non-differentiated colonies were isolated and further expanded.  Stemness of the isolated clones was confirmed in situ using Anti-TRA-1-60-Vio488 antibody (Miltenyi).  Patient-derived iPSC lines are grown on Geltrex (Gibco)-coated plates in Essential 8 medium (Gibco).   Following each passage, differentiated cells were eliminated under microscope, and stemness of the maintained cells were constantly monitored with immune-fluorescent analysis with anti-TRA-1-60-Vio488  and other stem cell specific antibodies. Genomic DNA of iPSCs was extracted using DNeasy Blood and Tissue kits (Qiagen). DNA library was prepared using TruSeq PCR-Free Kit (Illumina, San Diego, CA) according to the manufactural protocol.