Cell Biiology Core

核心B细胞生物学核心(核心B)的目标是通过提供来自特定年龄的小鼠的睾丸、卵巢和丰富的生殖细胞来实现计划的目标，以支持B Petkov、C Hibbs和D Handel项目中的ChlP-seq实验；C Hibbs和D Handel项目中的RNA-seq实验；以及D Handel项目中的RNA和蛋白质表达分析。Core B将利用精心控制的方法从幼年睾丸中获取生殖细胞，用于分析高度同步的第一波精子发生的前沿。此外，Core B将使用单位重力下的细胞沉淀技术，从成人睾丸中浓缩粗线型精母细胞和圆形精子FID。核心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将在亨德尔博士的指导下进行，亨德尔博士在减数分裂和男性生殖细胞生物学研究方面经验丰富，在分离小鼠精母细胞方面拥有专业知识。总而言之，核心B的功能确保所有项目都将拥有高质量和一致的实验材料，以实现个别项目目标以及集体实现计划的总体目标。155细胞生物学核心(Handel)Paigen，Kenneth的具体目标细胞生物学核心(核心B)将支持该计划的细胞和分子需求。这一目标将确保所有项目中的生殖细胞和生殖细胞提取物的高质量和一致的制备，从而使该计划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，将这些职能集中到一个核心中，可以提高成本和功能的效率。核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(D Handel项目)、RNA-seq(C Hibbs和D Handel项目)和ChlP-seq(B Petkov、C Hibbs和D Handel项目)。细胞生物学核心的目标是：具体目标1。核心将从小鼠身上回收性腺，并从幼年和成熟小鼠身上制备丰富的生殖细胞，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，用于该方案的表达分析和RNA-SEQ实验。具体目标3.核心将在性腺或生殖细胞上进行染色质免疫沉淀，以支持该计划的ChlP-SEQ实验。总体而言，这些目标确保所有研究人员将拥有必要的实验材料，以实现他们各自的项目目标以及共同实现项目的总体目标。

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.