SleepingBeautytransposonmutagenesisinratspermatogonialstemcells
Sleeping Beauty transposon mutagenesis in rat spermatogonial stem cellsZoltán Ivics,1, 2 Zsuzsanna Izsvák,1, 2 Gerardo Medrano,3, 4 Karen M Chapman3, 4 & F Kent Hamra3, 4Nature Protocols Volume: 6, Pages: 1521–1535 Year published: (2011) DOI: doi:10.1038/nprot.2011.378 Published online 08 September 2011 AbstractWe describe an experimental approach for generating mutant alleles in rat spermatogonial stem......阅读全文
基因敲除技术概述（四）
[13]。2.3.2 RNAi基因敲除的优点及应用①.比用同源重组法更加简便，周期大大缩短。②.对于哺乳动物，如对于一些敲除后小鼠在胚胎时就会死亡的基因，可以在体外培养的细胞中利用RNAi技术研究它的功能。③.由于RNAi能高效特异的阻断基因的表达，它成为研究信号传导通路的良好工具。④.RNAi还被
Fluorescent－Staining－of－Cells
1. Fluorescent phalloidin in methanol. Phallacidin does not work as well. Dilute 10 ul 330 nM stock into 500 ul PBS for each large coverslip.2. PB
Freezing－and－Thawing－cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Electroporation－of－ES－cells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for
Isolation－of－papillary－cells
Isolation of renal papillary cells1. For isolation of papillary cells, kidneys were harvested and kept in HBSS containing 15 mM HEPES, penicillin/
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Transfecting－Suspension－Cells
实验概要将转移基因整合到细胞染色体DNA上,形成稳定表达转移基因的细胞系。 实验原理 细胞转染技术是目前广泛应用于病毒基因结构与功能以及基因调控等的研究。细胞转染可分为短暂转染和稳定(或永久) 转染两种。在短暂转染中,被转染基因并不整合至细胞染色体中,因而不能随细胞分裂而传代。转入病毒基因的转
KARYOTYPING－ES－CELLS
An actively growing culture of cells is required, i e 2 - 3 d ES cell culture. The total number of cells needs to be between 106 - 107 cells.N B Read
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important.Prepare a Pasteur pip
Preparing－cells－and...
实验概要The method provides a protocol and tips for BrdU staining in tissue sections.Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic
Subculturing－Adherent－Cells
实验概要The following protocol describes a general procedure for subculturing adherent mammalian cells in culture.主要试剂1. Complete growth medium, pre-warme
Isolation－of－papillary－cells
实验概要This protocols provides a general protocol for isolation of papillary cells.实验步骤Isolation of renal papillary cells1. For isolation of papillary c
Immunofluorescence－Labeling－of－Cells
实验概要Antibodies are an important tool for demonstrating both the presence and the subcellular localization of an antigen. Cell staining is a very ver
研究揭示RNA甲基化修饰调控哺乳动物细胞微环境维持机制
近期，中国科学院西北生态环境资源研究院西北高原生物研究所研究员杨其恩课题组以小鼠为模型，揭示RNA甲基化修饰调控哺乳动物精原干细胞微环境维持的新机制。 成体干细胞命运决定受到特殊微环境调控，在大多数组织中，微环境的形成和维持机制并不明确。精原干细胞是一类经典的成体干细胞，是哺乳动物精子发生的基
关于CRISPRCAS9介导的基因编辑细胞株的一些经验总结
最近小编在应用CRISPR/CAS9作为工具制备基因编辑细胞株，有一些阶段性总结和设想，在这里贴出来，大家可以互相探讨一下。首先要描述一下“基因编辑细胞株”的具体要求：（1）“基因编辑”是指在特定位点引入期望的碱基修饰。具体过程为，通过CRISPR/CAS9在待编辑位点（靶位点）附近引入基因组DNA
关于CRISPRCAS9介导的基因编辑细胞株的一些经验总结
最近小编在应用CRISPR/CAS9作为工具制备基因编辑细胞株，有一些阶段性总结和设想，在这里贴出来，大家可以互相探讨一下。 首先要描述一下“基因编辑细胞株”的具体要求： （1）“基因编辑”是指在特定位点引入期望的碱基修饰。 具体过程为，通过CRISPR/CAS9在待编辑位点（
AbC™－AntiRat/Hamster－Bead－Kit
实验概要The AbC™ anti-Rat/Hamster Bead Kit provides a consistent, accurate, and simple-to-use technique for the setting of flow cytometry compensation
AbC™－AntiRat/Hamster－Bead－Kit
实验概要The AbC™ anti-Rat/Hamster Bead Kit provides a consistent, accurate, and simple-to-use technique for the setting of flow cytometry compensation
stem－cell－culture－protocol
实验概要stem cell culture protocol主要试剂cell culture supplies and reagentssEnvironment: cell culture requires a sterile environment, so it needs a separat
stem是什么仪器？
扫描透射电子显微镜(scanning transmission electron microscopy，STEM)既有透射电子显微镜又有扫描电子显微镜的显微镜。STEM用电子束在样品的表面扫描，通过电子穿透样品成像。STEM技术要求较高，要非常高的真空度，并且电子学系统比TEM和SEM都要复杂。
胚胎干细胞不同阶段的marker表
Digging through the literature, I've compiled a list of possible markers for various cell types, which I'll publish here for the benefit of an
Intestinal－intraepithelial－lymphocytes
Intestinal intraepithelial lymphocytes Intestinal intraepithelial lymphocytes (IELs) are mostly T cells, which are continuously associated with gut
FACS－Analysis－of－ES－Cells
Isolate cells and dissociate to single cell suspension (can use Gibco Cell Dissociation Buffer, Accutase or Trypsin)Wash with 10% FBS/DMEM:F12For surf
Isolation－of－rodent－pancreatic－β－cells
1. Adult rats weighing 250-350g were anesthetized, sacrificed and immediately used for pancreas sampling.2. Rat islets were isolated from male wistar
Freezing－cells－in－liquid－nitrogen
Take off MediaTrypsinate with 1ml x2 Dulbecco A trypsinAdd 7ml MediaPipette up and down to distribute cells throughout media (i.e. not clumped togethe
Decontamination－of－cells－from－the－yeast
I Destroy yeast1. Aspirate medium and wash cell in PBS.2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.3. In
Selection－of－Transfected－Suspension－Cells
Contributor: Suprya JayadevDate: December 13, 19941) Transfect cells.2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g.
Routine－Culturing－of－ES－Cells
Cell are normally passaged every 2-3 days, this is important to avoid differentiation.Signs of differentiation are:-i) colonies are surrounded by flat
Isolation－of－lymphatic－endothelial－cells
Dermal Cell Suspensions 1. Dermatomed 0.8-mm split-thickness skin was obtained from adult healthy individuals undergoing elective surgery. 2
RNAse－A－Treatment－of－Mouse－Cells
IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into