Transcriptome analysis of meiotic prophase spermatocytes in Zfp541 +/- and Zfp541 KO mice

ZFP541 is one of the candidates for a transcriptional regulator during mouse meiotic prophase. In order to address the role of ZFP541, transcriptomes of meiotic prophase-enriched population were compared between Zfp541 +/- and Zfp541 KO testes by RNA-seq. The meiotic prophase-enriched population was isolated from Zfp541 +/- and Zfp541 KO testes at postnatal day 18 (P18), on the Rec8-3FH-GFP KI background by fluorescent sorting of GFP positive cells. GFP positive cells from Zfp541 KO and the control heterozygous on the Rec8-3xFLAG-HA-p2A-GFP knock-in background testis were sorted using SH800S cell sorter. (RNA-seq for 1 testis from Zfp541 +/-: Rec8-3FH-GFP KI hetero, 1 testis from Zfp541 +/-: Rec8-3FH-GFP KI homo, 2 individual testes from Zfp541 KO: Rec8-3FH-GFP KI homo). Total RNAs were prepared by Trizol and quality of total RNA was confirmed by BioAnalyzer 2100 (RIN > 8). Library DNAs were prepared according to the Illumina Truseq protocol using Truseq Standard mRNA LT Sample Prep Kit (Illumina) and sequenced by Illumina NextSeq 500 (Illumina) using Nextseq 500/550 High Output v2.5 Kit (Illumina) to obtain single end 75 nt reads.