RNA-Seq (Ribo- library) of Mus musculus: newborn retina
Retinas from mice were dissected in RNAlater (Qiagen) and total RNA was extracted using Qiagen RNeasy Mini Kit (Qiagen). Samples were treated with DNAse (TURBO DNA-free, Thermo Fisher Scientific) following manufacturers protocol and the absence of genomic DNA contamination was confirmed by RT-PCR and was found to be <3 pg/ml. Library preparation and sequencing was performed at the Next Generation Sequencing Core from Scripps Research Institute (San Diego, CA). Strand-specific libraries were generated by rRNA depletion using the NEB Next Ultra Directional RNA Library Prep Kit for Illumina (New England BioLabs), following manufacturers protocol. Purified libraries were quantitated using the Qubit quantitation platform (Invitrogen) and sized using RNA Nano Chip (Agilent). Single-read sequencing was performed on the Illumina HiSeq 2000 platform and demultiplexed based on index sequences.
SUB15723501
R12_2