BioProject

Purpose: To test the effect of AHCY O-GlcNAcylation on E14.1cell pluripotency and The goals of this study are to compare gene expression profiles of the WT AHCY rescue cells and the T136A AHCY rescue cells. More...

Purpose: To test the effect of AHCY O-GlcNAcylation on E14.1cell pluripotency and The goals of this study are to compare gene expression profiles of the WT AHCY rescue cells and the T136A AHCY rescue cells. Method:Total RNA was Trizol-extracted and purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in NEB. cDNA was synthesized using random hexamer primer and RNase H. The library fragments were purified with QiaQuick PCR kits and elution with EB buffer, then terminal repair A-tailing and adapter added were implemented. The clustering of the index-coded samples was performed on a cBot cluster generation system using HiSeq PE Cluster Kit v4-cBot-HS (Illumina) according to the manufacturer’s instructions. After cluster generation, the libraries were sequenced on an Illumina platform and 150 bp paired-end reads were generated. Results:Significant alterations in gene expression levels were observed, with 1351 genes upregulated and 979 genes downregulated in the T136A AHCY rescue cells Overall design: the mRNA profiles of WT AHCY E14.1 cells and T136A AHCY E14.1 cells were generated by deep sequencing using Illumina GAIIx. Less...