RNAFISHonculturedcellsininterphase2
Post-labeling DNA processing and purificationQiagen PCR clean up to get rid of unused oligonucleotides;Add 20µl cot1DNA, 10µl ssDNA to compete for repetitive elements;Precipitate the probe with 1/10 V 3 M NaOAc pH5.2, 2.5 V 100% ethanol at -80°C for 30 minutes;Spin down pellet at 4°C for 20 minutes;Wash pellet with 70% ethanol;Resuspend pellet in 80µl hybridization mix.The probe is now ready to be used. It can be sto......阅读全文
Peripheral－blood－“endothelial－progenitor－cells”
EPC Isolation and Characterization1. EPCs were obtained by isolating mononuclear cells using Ficoll density-gradient centrifugation of human blood buf
Differentiating－Neural－Stem－Cells－into－Neurons－and－Glial－Cells
实验概要The protocols in this section describe the steps involved in differentiating neural stem cells (NSC) to neurons, astrocytes, and oligodendrocyte
Isolation－of－human－primary－gastric－mucosa－epithelial－cells
1. A piece of gastric mucosa (∼3 cm2) was obtained from the normal appearing mucosa of the stomach at surgery. 2. The patients were proven to be no H.
DNA微序列技术
· Protocols for Making Drosophila Arrays (Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,
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 fila
Effect－of－ethanol－on－development－of－Danio－reriro
ObjectiveThe objective of the experiment is to determine the effects of ethanol exposure on the embryonic development of zebrafish through observation
mpulsive－Pressurization－of－Neuronal－Cells－for－Traumatic－Brain－Injury－Study
实验概要 A novel impulsive cell pressurization experiment has been developed using a Kolsky bar device to investigate blast-induced traumatic brain in
Growing－cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irresp
Trypsinizing－cells
There are many procedures with which to trypsinize cells. All include washing the cell monolayer with TD, or in rare cases, with VE. This removes ser
Lyophilizing－Cells
Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized. Incubate the culture at 37°C with vig
Lyophilizing－Cells
Lyophilizing Cells Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized. Incubate the c
Freezing－Cells
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
Isolation－of－human－prostatic－smooth－muscle－cells
Human prostate tissues1.Human hyperplastic prostates were obtained during surgery from four men through transurethral resection of the prostate. 2.All
Production－of－Recombinant－Proteins－in－SuspensionCultured－Plant－Cells
Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, toda
细胞组分和细胞器——染色体
Chromosomal DNA Prep : cultured cells/tissue samples (Mike A Dyer)This protocol was developed for cultured cells but should be appropriate for dissoci
FISH和PRINS技术
一、荧光原位杂交（FISH） 是一种简单、敏感、而容易操作的方法，结果迅速，并能在同一标本上检测多个不同的基因，可应用于细胞培养、染色体分裂像、冰冻切片和石蜡切片。 FISH技术是利用特异的DNA探针，标记了生物素，地高辛、或荧光素，对检测细胞进行DNA-DNA原位杂交，并用荧光法显示。 FISH
FISH－技术基本实验
基本方案 实验方法原理 试剂、试剂盒 70%乙醇 SSC
FISH－技术基本实验
实验方法原理试剂、试剂盒70%乙醇SSC磷酸盐缓冲溶液（PBS)Hemo-De清理试剂蛋白酶溶液仪器、耗材水浴锅增湿器玻片实验步骤一、制备培养的中期细胞标本1.在一个独立小室里放置一个水浴锅和一台增湿器。湿度应大约50%，如果房间湿度计显示低于45%，则应使用加湿器。2.预热水浴锅至67℃±2℃，在
FISH－技术基本实验
实验方法原理 试剂、试剂盒 70%乙醇SSC磷酸盐缓冲溶液（PBS)Hemo-De清理试剂蛋白酶溶液仪器、耗材 水浴锅增湿器玻片实验步骤 一、制备培养的中期细胞标本1.在一个独立小室里放置一个水浴锅和一台增湿器。湿度应大约50%，如果房间湿度计显示低于45%，则应使用加湿器。2.预热水浴锅至67℃±
Timing－of－Cycles
Materials Monolayer cultures grown in 75 mm culture flasks (Cells from Exercise 11.4 may be used, or cultures of tetrahymena, yeast, or algae ma
Combined－Flow－Cytometric－Measurement－of－Two－CellSurface－Antigens
INTRODUCTION Flow cytometry is frequently used to assess nucleic acid content in individual cells. Based on DNA content alone, however, cells in the
显微镜技术——电子显微技术
The Transmission Electron Microscope (TEM) (HEI)An explanation of how the TEM works. TEM Specimen Preparation (HEI) Serial Sectioning (Walter Steffe
Primary－brain－cell－isolation－and－culture
1. Cerebella were removed from 7-day-old mice and passed through Nitex nylon netting (80 μm pore size) into primary cell system containing 20% (v/
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/
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.
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 Re
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 syntheti
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 cells 1. For isolation of pa