Abstract：
Chromatin undergoes extensive remodeling during meiosis, leading to specific patterns of gene expression and chromosome organization, which ultimately controls fundamental meiotic processes such as recombination and homologous chromosome associations. Recent game-changing advances have been made by analysis of chromatin binding sites of meiotic specific proteins genome-wide in mouse spermatocytes. However, further progress is still highly dependent on the reliable isolation of sufficient quantities of spermatocytes at specific stages of prophase I. Here, we describe a combination of methodologies we adapted for rapid and reliable isolation of synchronized fixed mouse spermatocytes. We show that chromatin isolated from these cells can be used to study chromatin-binding sites by ChIP-seq. High-quality data we obtained from INO80 ChIP-seq in zygotene cells was used for functional analysis of chromatin-binding sites.
摘要：
减数分裂过程中染色质经历广泛的重塑，导致基因表达和染色体组织的特定模式，它最终控制基本的减数分裂过程，如重组和同源染色体关联。通过分析小鼠精母细胞全基因组减数分裂特异性蛋白质的染色质结合位点，已经取得了最新的改变游戏规则的进展。然而,进一步的进展仍然高度依赖于在I期前期的特定阶段可靠地分离足够数量的精母细胞，我们描述了我们适用于快速，可靠地分离同步固定的小鼠精母细胞的方法组合。我们表明，从这些细胞中分离的染色质可用于通过ChIP-seq研究染色质结合位点。我们从合子细胞中的INO80ChIP-seq获得的高质量数据用于染色质结合位点的功能分析。
