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TissueCultureMethods3

REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should maintain his own cells throughout the course of the experiment. These cells should be monitored daily for morphology and growth characteristics, fed every 2 to 3 days, and subcultured when necessary. A minimum of two 25 cm2 flasks should be carri......阅读全文

Testing for Mycoplasma by Indirect DNA Stain (Hoechst 33258 stain)

AimDNA staining methods such as Hoechst staining techniques are quick with results available within 24 hours, which compares favorably with 4 weeks fo

细胞培养——培养基和试剂

·         Cell Culture Media and Solutions (Donis-Keller lab)·      &n

细胞培养——细胞保藏

Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank    Maste

BrdU Labeling Protocol

实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic

细胞培养常规操作

常规操作(主要内容如下)·         Aseptic Technique·         Culture Ves

Fibroblast Cell Systems-2

Handling Precautions Normal human cells are fragile, and require special handling:Upon receipt, immediately store cryopreserved cells i

Mouse keratinocyte cultures

PRIMARY MOUSE KERATINOCYTE CULTURESIsolation of epidermal keratinocytes from neonatal mice is based on the protocol of Dlugosz et al., Methods En

Methods for the Detection of D-Amino-Acid Oxidase-1

AbstractFour methods (an enzyme activity assay, western blotting, RT-PCR, and northern hybridization) to detect the enzyme D-amino-acid oxidase are de

Isolation of human primary ovarian surface epithelial cells

1. Surface epithelial cells from normal ovaries from surgical residual specimen were isolated using standard and Institutional Review (Refere

Fibroblast Cell Systems-4

3. If your result falls into any quadrant other than the "High Yield-High Viability" quadrant, refer to Appendix D, Improving Cell Yield and

E.Z.N.A.® Total RNA Midi Kit Protocol for Eukaryotic Cells and Tissues

实验概要E.Z.N.A.®  Total RNA Midiprep Kit provides a rapid and easy method for the  isolation of up to 600 ug of total RNA from cultured eukaryo

Embryonic limb bud culture in media

Early in embryonic development, the region of the chick embryo which is determined to form a limb first differentiates from the rest of the embryo1. T

慢病毒转染肝细胞方法

Lentivirus Transduction of Hematopoietic CellsMing-Jie Li and John J. RossiDivision of Molecular Biology, Beckman Research Institute of the City of Ho

核蛋白基因组指纹技术

Fine Mapping of Genomic Targets of Nuclear Proteins in Cultured CellsAchim Breiling and Valerio OrlandoDulbecco Telethon Institute, Institute of Genet

哺乳动物RNAi技术 Mammalian RNA Interference

Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide

定量RT-PCR (Quantitative RT-PCR)

Application: Quantitative RT-PCR is used to quantify mRNA in both relative and absolute terms. It can be applied for the quantification of m

显微镜技术——荧光显微技术

Immunofluorescencc Microscopy of tissue culture cells (Microscopy and Electronic Imaging Lab)These methods are written for direct staining of fil

Culturing Human Neural Stem Cells

实验概要Neural  stem cells (NSC) are valuable resources because of their ability to  differentiate into neurons and glial cells with application

In Planta Transformation of Arabidopsis

实验概要        A breakthrough in Arabidopsis research was the invention ofthe vacuum-infiltration procedure, a simple and reliabl

Crystallization Trials

MaterialsMilipore filter type HA 0.45 micronCulture plates (Linbro model 76-033-05)Protein in DDW or HEPES buffer (10 mg/mL)Vacuum greaseSyringe with

Immunofluorescence Microscopy of tissue culture cells

Immunofluorescence Microscopy of tissue culture cellsThese methods are written for direct staining of filamentous actin with bodipy FL-phallicidin and

Tissue Culture Methods-2

IV. MAINTENANCECultures should be examined daily, observing the morphology, the color of the medium and the density of the cells. A tissue culture log

Aimee''''s non-Radioactive EMSA Protocol

NUCLEAR EXTRACTION OF TISSUE CELLSThis procedure was developed for HNEK cells grown in KGM, from Clonetics.  All steps are performed on ice, with

通过细胞受体代谢生物素化进行图像分析

Metabolic biotinylation of mammalian cell receptors for imagingBakhos A. Tannous , btannous@hms.harvard.edu, Massachusetts General Hospital and Harvar

Hints and precautions for the care, feeding and breeding of Neurospora-2

9. Spontaneous germination. Some genotypes result in spontaneous ascospore germination (for example, the unpigmented ascospores of per-1 Type I). A si

Preparation of nucleic acid probes

Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s

Isolation and Culture of Multipotent Stem Cells from Human Bone Morrow

Bone marrow contains three types of stem cells:1. Hematopoietic stem cells give rise to the three classes of blood cells that are found in the

Beta-gal staining of eukaryotic cells in vitro

(Modification of methods of Dr. Seong-Seng Tan and Promega's "Protocols and Applications Guide")Cells previously transfected with a lac

活细胞提取及应用——单个细胞级别的活细胞提取

由于细胞异质性的存在,单细胞层面的分析就变得十分重要。目前对于单细胞分析的方法主要还是通过化学、生物学的方法进行裂解后,提取内容物进行分析,然而这种方法往往会对样本造成一些损伤。直接提取活细胞具有诸多优点,但是操作苦难。如今一种全新使用FluidFM科技的技术新报道有望提供一种活细胞提取新型的简易方

DNA转染

DNA转染·         Transfection of Mammalian Cells Using Lipofectamine (LTI)·     &n