Illumina HiSeq 2000 sequencing; Small RNA profiling of TUT4 and TUT7-deficient mouse pachytene-diplotene cells.
Small RNA profiling of TUT4 and TUT7-deficient mouse pachytene-diplotene cells.
Testes from adult mice were dissected, decapsulated and digested with collagenase and trypsin under gentle agitation at 32°C followed by pipetting to obtain a single-cell suspension. Cells were then stained with Hoechst 33342 and propidium iodide (PI). Pachytene-diplotene cells were FACS sorted according to their characteristic blue/red Hoechst fluorescence profile and GFP-high expression. Total RNA was extracted using QIAzol lysis reagent (Qiagen) according to manufacturer's instructions. Libraries were prepared using the NEBNext Multiplex Small RNA Library Prep Set for Illumina following manufacturer's instructions
Experimental Factor: genotype
Ythdf2 +/GFP-Fl