GSM5165274: Q768_T1ND; Mus musculus; RNA-Seq
Mouse islets were digested from pancreatas and purified by Histopague gradient centrifugation. The isolated islets were trypsized and sorted through flow cytometer for  mCherry+ cells (β cells). The RNA extraction was performed on sorted cells via TRIzol phenol-chloroform or Qiagen silica gel spin column. Ultra-low RNA input libraries are generated using the Takara “SMART-Seq v4 Low Input RNA Kit for Sequencing” (Takara, Mountain View, CA, USA) for cDNA synthesis and the Illumina NexteraXT DNA Library Preparation (Illumina, SanDiego, CA, USA) kit for cDNA dual indexing.  Full length cDNA fragments are generated from 1-10ng total RNA by the Takara SMART (Switching Mechanism at 5' End of RNA Template) technology.  cDNA fragments are fragmented  and dual indexed in a single step by taking advantage of the Nextera kit's simultaneous transposon and tagmentation step. Libraries were standardized to 2nM.