ElectroporationofEScells
Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for 1-2 electroporations.Procedure1. Change medium on ES cells 3-4 hours prior to electroporation2. Gelatinize 10 cm plates, then add 10 ml medium to each.3. Place them in a 37 0C incubator until they are required.4. Switch on the electroporation apparatus.5. Harvest ES cel......阅读全文
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
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 co
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
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: Tryp
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
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
Screen-ES-cells-by-Southern-Blot
Digest DNA in 96-well plate To each well add: 4ul 10Xbuffer 4ul Enzyme 0.4ul Spermidine(0.4M) 31.6ul H2O 37‡C 19h, then add 4ul loading dye to each w
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: Try
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 colon
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
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
Human-Embryonic-Stem-(ES)-Cell-Protocols—Splitting-Human-ES-cells-onMatrige
based on splitting onto on plateWarm collagenase media to 37°C in a water bath.Aspirate media off of cell culture plate.Add the following amount of co
Human-Embryonic-Stem-(ES)-Cell-Protocols——Splitting-Human-ES-cells-on-MEFs
based on splitting onto one plateWarm collagenase IV split media to 37 °C in a water bath.Aspirate media off of cell culture plate.Add the following a
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 (
胚胎干细胞培养技术大全
MEDIA AND SOLUTIONS REQUIRED FOR ROUTINE ES CELL CULTURE Routine Culturing of ES Cells ISOLATION OF PRIMARY MOUSE EMBRYO FIBROBLASTS MITOM
ES-Cell-Culture-and-Manipulation
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/stre
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
Competent-agro-prep-for-electroporation
day 1 1. Start 75 mL overnight cultures of agro (strain GV3101 C58C1 Rifr pMP90 Gmr, Koncz & Schell) in YEP in 250 mL baffle flasks. 2. Grow at
Streptomyces:Protocols/Transformation-by-Electroporation
Description Transform E.coli cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into E.coli).Approx. Duration:Prep
Growing-feederindependent-embryonic-stem-cells§
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
SingleCell-Electroporation-in-Xenopus
Single-Cell Electroporation in Xenopus Xue Feng Liu and Kurt Haas INTRODUCTION Single-cell electroporation (SCE) is a versatile technique for del
Transformation-of-E.-coli-by-Electroporation
实验概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc
Culture-of-BEND-Cells-(Bovine-Endometrial-Cells)
Culture of BEND Cells (Bovine Endometrial Cells) Charles E. Krininger, III and Peter J. Hansen Dept. of Animal Sciences, University of Florida Thi
Electrotransformation-of-Lactobacillus-Spp.
OverviewGeneral guidelines for the electro-transformation of Lactobacillus sake as described by Berthier et al. and as used by Alegre et al. with Lact
Bacterial-transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
革兰氏阳性菌的电转化方案(英文)
Transformation of Gram-Positive Bacteriaan adaptation from Chang, D., Chassy, B., Saunders, J., Sowers, A. 1992.Guide to Electroporation and Electrofu
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
酵母转化
· Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation· Yeast Transformation (Breeden Lab)LiAc method· Large-Scale Y