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......閱讀全文
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
胚胎干細胞培養
Media and Solution required for ES Cell Culture?(Bowtell Lab)???Routine Culturing of ES Cells?(Bowtell Lab)??Routine Splitting and freezing of cells?(
胚胎干細胞培養技術大全
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
轉基因——基因標靶
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
轉基因
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
實驗概要? ? ? ? Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low?temperature, and
DNA轉化
DNA轉化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
革蘭氏陽性菌的電轉化方案（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
實驗概要The protocols in ?this section describe the steps involved in differentiating neural stem ?cells (NSC) to neurons, astrocytes, and oligodendrocyte
酵母轉化
·?????????Yeast Transformation?(Gietz Lab)LiAc/SS-DNA/PEG Transformation·?????????Yeast Transformation?(Breeden Lab)LiAc method·?????????Large-Scale Y