Next Generation Sequencing of p53 KO and Utx/p53 DKO Hematopoietic Stem Cells Transcriptomes

Hematopoitic stem cells from Utx/p53 DKO mice showed increased self-renewal potential and predisposition of differentiation to myeloid lineage compared with those from p53 KO mice. To figure out what mechanism leads to the increased self-renewal, we compared the transcriptional profiles using RNA-sequencing of FACS sorted LSK (Lin-Ckit+Sca1+) cells, which contains the hematopoietic stem cells. Overall design: Examination of differential genes expression in 2 genotipic hematopoitic stem cells LSK cells were sorted directly into TRIzol (Invitrogen),then column-purified with Direct-zol RNA MiniPrep Kit (Zymo research). 50ng total RNA was used for preparation of cDNA sequencing library. Sample libraries were pooled for 100bp double read sequencing on Illumina HiSeq2500 resulting in 50 million reads per sample.