Cell Biiology Core

细胞生物学核心（CORE B）的目标是通过提供来自特定年龄小鼠的睾丸、卵巢和富集生殖细胞来支持项目B Petkov、C Hibbs和D Handel中的chp -seq实验，从而实现该计划的目标；项目C Hibbs和D Handel的RNA-seq实验以及Project D Handel的RNA和蛋白表达分析。核心B将利用精心控制的方法从少年睾丸中获得生殖细胞，用于高度同步的第一波精子发生的前沿分析。此外，Core B将使用单位重力下的细胞沉降技术，从成年睾丸中富集粗线精细胞和圆形精子。Core将为项目中提出的实验执行RNA纯化和染色质免疫沉淀的初始步骤。核心B将由亨德尔博士指导，他在减数分裂和男性生殖细胞生物学的研究方面经验丰富，并在分离小鼠精细胞方面具有专长。综上所述，核心B的功能确保所有项目将拥有高质量和一致的实验材料，以实现单个项目目标以及共同实现计划的总体目标。155细胞生物学核心（Handel Paigen, Kenneth SPECIFIC AIMS）细胞生物学核心（核心B）将支持计划的细胞和分子需求。这一目标将确保所有项目中生殖细胞和生殖细胞提取物制备的高质量和一致性，从而使项目中的实验结果具有直接可比性。此外，在经验丰富的领导者的指导下，将这些功能集中到一个核心中，可以提高成本和功能的效率。该中心将为利用性腺或生殖细胞的实验提供材料，包括表达分析（项目D Handel）、rna测序（项目C Hibbs和D Handel）和chp测序（项目B Petkov、C Hibbs和D Handel）。细胞生物学核心课程的目标是：该中心将从小鼠中提取性腺，并从幼年和成年小鼠中制备富集的生殖细胞，用于该计划的实验。具体目标2。核心将从性腺和/或生殖细胞中提取RNA和蛋白质，用于项目的表达分析和RNA-seq实验。具体目标3。核心将对性腺或生殖细胞进行染色质免疫沉淀，以支持该计划的ChlP-seq实验。总的来说，这些目标确保所有研究人员都有必要的实验材料来实现他们个人的项目目标以及集体实现该计划的总体目标。

CORE B The objective of the Cell Biology Core (Core B) is to enable goals of the Program by providing testes, ovaries and enriched germ cells from mice of specific ages to support ChlP-seq experiments in Projects B Petkov, C Hibbs and D Handel; RNA-seq experiments in Projects C Hibbs and D Handel; and RNA and protein expression analyses in Project D Handel. Core B will utilize carefully controlled methods to obtain germ cells from juvenile testes for assays that follow the leading edge of the highly synchronized first wave of spermatogenesis. Additionally, Core B will use a technique of cell sedimentation at unit gravity to enrich pachytene spermatocytes and round sperm fids from adult testes. The Core will perform initial steps of RNA purification and chromatin-immunoprecipitation for experiments proposed in the Projects. Core B will be under the direction of Dr. Handel, who is highly experienced in investigation of meiosis and male germ-cell biology, and has expertise in isolation of mouse spermatocytes. Taken together, the functions of Core B ensure that all Projects will have high-quality and consistent experimental materials to achieve the individual Project goals as well as to collectively achieve the overall goals of the Program. 155 Cell Biology Core (Handel) Paigen, Kenneth SPECIFIC AIMS The Cell Biology Core (Core B) will support cellular and molecular needs of the Program. This goal will ensure high-quality and consistent preparations of germ cells and germ-cell extracts across all Projects, so that experimental results in the Program will be directly comparable. Moreover, centralizing these functions into a Core, under the direction of an experienced leader, provides efficiencies of cost and function. The Core will provide material for experiments that utilize gonads or germ cells, including expression analyses (Project D Handel), RNA-seq (Projects C Hibbs and D Handel), and ChlP-seq (Projects B Petkov, C Hibbs and D Handel). The aims for the Cell Biology Core are: Specific Aim 1. The Core will retrieve gonads from mice and prepare enriched germ cells from juvenile and mature mice to be used in experiments of the Program. Specific Aim 2. The Core will prepare extracts of RNA and protein from gonads and/or germ cells to be used for expression analyses and RNA-seq experiments of the Program. Specific Aim 3. The Core will conduct chromatin-immunoprecipitation on gonads or germ cells to support ChlP-seq experiments of the Program. Overall, these aims ensure that all investigators will have the necessary experimental materials to achieve their individual Project goals as well as to collectively achieve the overall goals of the Program.