GSM3722564: mRNA_C1483Y_3; Mus musculus; RNA-Seq
RNA was extract using Trizol Libraries were prepared according to reference (Cho et al., Science 2015, Oct 2;350(6256):82-7. doi: 10.1126/science.aac7368). Libraries were prepared according to Illumina's instructions accompanying the TruSeq small RNA Kit (Cat#RS-200-0012). Briefly, Ribosome protected fragment (RPF) were purifed using a RNase I and Sephacryl S-400 column. mRNAs were heat fragmented. RPF and mRNAs were labeled with 32P using a T4 polynucleotide kinase. RPF and mRNAs were size selected usign Urea-PAGE. RPFs and mRNAs were end-repaired using antarctic phosphatase and T4 polynucleotide kinase. After purification, 3' adaptor and 5' adaptor ligated and purifed by UREA PAGE. After RT-PCR, libraries were sequenced using Hiseq 2000.