ELECTROPORATIONOFESCELLSANDISOLATIONOFH／RCLONES
Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold PBS.2. Take x2 Biorad curvettesA. 0.9 ml (107 cells)B. 0.167 ml cells (2 x 106 cells ) + 0.73 ml PBSTo A add 15 ug linearised plasmid DNA, IMMEDIATELY prior to electroporation..B no DNA3. Using a Biorad Gene Pulser, (McMahon, A.P., and Bradley, A. (1990) Cell 62 1073-10......阅读全文
ELECTROPORATION－OF－ES－CELLS－AND－ISOLATION－OF－H/R－CLONES
Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold
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
胚胎干细胞培养
Media and Solution required for ES Cell Culture (Bowtell Lab) Routine Culturing of ES Cells (Bowtell Lab) Routine Splitting and freezing of cells (
转基因——基因标靶
Gene Targeting Outline (University of Michigan Transgenic Animal Model Core)This is a brief outline of the steps necessary to produce mice with a muta
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
DNA－Isolation－From－BAC－－PAC－Clones
DNA Isolation From BAC & PAC Clones This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a
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
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
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/
Differentiate－ES－cells－into－glial－cells－and－neurons
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsiniz
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
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
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
Isolation－of－lymphatic－endothelial－cells
实验概要This protocols provides a general protocol for isolation of lymphatic endothelial cells.实验步骤Dermal Cell Suspensions1. Dermatomed 0.8-mm split-thic
Screen－ES－cells－by－Southern－Blot
Digest DNA in 96-well plateTo each well add:4ul 10Xbuffer4ul Enzyme0.4ul Spermidine(0.4M)31.6ul H2O37‡C 19h, then add 4ul loading dye to each well. Lo
Differentiate－ES－cells－into－cardiac－myocytes
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.____________________Day 1: Trypsini
Isolation,－Culture,－and－Differentiation－of－Progenitor－Cells
Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous SystemScott R. Hutton and Larysa H. Pevny1UNC Neuroscience Center
Isolation－of－Human－Pulmonary－Epithelial－Cells
Isolation of Human Pulmonary Epithelial Cells 1. Pulmonary epithelial cells from human lung tissue were isolated.2. Lungs were perfused with 10
Isolation－of－normal－mammary－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
Isolation－and－culture－of－porcine－muller－cells
1. Porcine eyes were harvested from pentobarbital-anesthetized animals, each weighing 30 to 40 pounds, and transported to the laboratory in ice-co
Isolation－of－human－endometrial－epithelial－cells
Tissue collection1. Endometrial biopsies were collected from women undergoing gynaecological procedures for benign conditions. 2. All women reported
Isolation－of－human－atrial－or－ventricular－cells
1. Human tissue was derived from atrial or ventricular biopsy specimens belonging to patients undergoing heart surgery.2. Isolated myocardial tissue w
Isolation－and－culture－of－pancreatic－stellate－cells
Pancreatic stellate cells (PaSCs or PSCs) are myofibroblast-like cells found in the areas of the pancreas that have exocrine function. PaSCs are
Isolation－of－human－prostatic－epithelial－cells
1. A small piece of tissue from each specimen was removed and minced. 2. The tissue was digested with collagenase overnight. 3. To remove the collagen
Human－Embryonic－Stem－(ES)－Cell－Protocols——Thawing－Human－ES－cells
Remove Human ES cells from liquid nitrogen storage tank. Fill out a freeze/thaw form.Thaw cryovial by gently swirling in waterbath until only a small
Human－Embryonic－Stem－(ES)－Cell－Protocols——Freezing－Human－ES－Cells
Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet, gently pipet and scrape colonies f
Differentiate－ES－cells－into－cystic－embryoid－bodies
Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.______________Day 1: Trypsinized th
Isolation－of－human－colonic－epithelial－cells－(EC)
1.Human colonic epithelial cells (EC) were isolated from freshly resected colonic surgical specimens obtained. 2.All diagnoses were confirmed by clini
Isolation－and－Culture－of－Human－Liver－Stem－Cells
Materials and Methods1. Human hepatocytes were isolated from fresh surgical specimens of patients undergoing hepatectomies. Healthy liver tissue (