title: GSM5252962: L1_RNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Murine spermatocytes were isolated by FACS as described previously (Gaysinskaya, Soh, van der Heijden, & Bortvin, 2014; Getun, Torres, & Bois, 2011) with modification. Wild-type or mutant mice of different ages were used for isolation of spermatocytes at different stages: 7-11 days old mice were used for isolating Sertoli, spermatogonia, and preleptotene cells, 2-week old mice for zygotene cells, 3- to 5-week-old mice for pachytene, diplotene, and Metaphase II cells, and 1-2 months old SUN1 W151R mutant mice for zygotene-like spermatocytes Total RNA was extracted from ~50k purified pre-leptotene, leptotene and zygotene spermatocytes by RNeasy Micro Kit (Qiagen, 74004) according to the manufacturer's instructions. The cDNA was synthesized and amplified using the Single Cell Full Length mRNA-Amplification Kit (Vazyme, N712). The RNA-seq libraries were constructed from the amplified cDNA using the TruePrep DNA Library Prep Kit V2 for Illumina® (Vazyme, TD503). The PCR and size-selection of RNA-seq libraries were conducted as described in Hi-C library construction. The RNA-Seq libraries were sequenced on Illumina Nova Seq 6000 platform at PE150 mode.
attributes.tag: GEO Accession
attributes.value: GSM5252962