ChIP-Seq experiment to analyse Zip1 localisation in the Ctf19c mutants during meiotic prophase.. Saccharomyces cerevisiae

The goal of this experiment is to confirm whether Ctf19c mutants affect Zip1 localisation during meiotic prophase. To do so, diploid S. cerevisiae SK1 cells were constructed containing ndt80-d to arrest cells in prophase. Cells were allowed to sporulate in sporulation media for 5 hours after which formaldehyde was added to the cultures and cells were fixed for 2hr. Samples were then processed according to the standard lab ChIP-Seq protocol (with 3 times 30cycles of 30sec of sonication) Overall design: Diploid S. cerevisiae SK1 cells were constructed containing ndt80-d to arrest cells in prophase. Cells were allowed to sporulate in sporulation media for 5 hours after which formaldehyde was added to the cultures and cells were fixed for 2hr. Samples were then processed according to the standard lab ChIP-Seq protocol (with 3 times 30cycles of 30sec of sonication)

本实验旨在验证Ctf19复合物（Ctf19c）突变体是否会影响减数分裂前期Zip1的定位。为此，我们构建了携带ndt80-d以将细胞阻滞于减数分裂前期的二倍体酿酒酵母（S. cerevisiae）SK1细胞。将细胞接种于孢子形成培养基中诱导孢子发生，培养5小时后向培养液中加入甲醛，固定细胞2小时。随后按照实验室标准染色质免疫共沉淀测序（ChIP-Seq）流程处理样本，超声破碎参数为3轮、每轮30次循环、每次30秒。整体实验设计：构建携带ndt80-d以将细胞阻滞于减数分裂前期的二倍体酿酒酵母（S. cerevisiae）SK1细胞。将细胞接种于孢子形成培养基中诱导孢子发生，培养5小时后向培养液中加入甲醛，固定细胞2小时。随后按照实验室标准染色质免疫共沉淀测序（ChIP-Seq）流程处理样本，超声破碎参数为3轮、每轮30次循环、每次30秒。