Illumina HiSeq 2000 paired end sequencing; RNA-seq Analysis of Chromatin Accessibility in p53 Deficient Spermatogonial Stem Cells for High Frequency Transformation into Pluripotent State
RNA-seq Analysis of Chromatin Accessibility in p53 Deficient Spermatogonial Stem Cells for High Frequency Transformation into Pluripotent State
SSCs were isolated from 5-day old p53+/+ or p53-/- mice testes using two-step enzymatic digestion and purified by FACS.The THY1+;c-kit- fraction was collected for centrifugation and collection. Total RNA was extracted using Trizol (Ambion Life Technologies) according to the Ambion standard RNA isolation procedure. mRNA was purified using the NEBNext Poly (A) mRNA Magnetic Isolation Beads (NEB, USA). Then the mRNA library was constructed with a NEBNext Ultra Directional RNA Library Prep Kit for Illumina (E7420S/L, NEB).