ProtocolforCellFusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, they should be centrifuged at a 64.4 xg on the IEC clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......阅读全文
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. Irrespe
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
Plastic－dishes－for－growing－cells
There are two kinds of dishes used to grow tissue culture cells.Those that are designed for adherent cells have been treated chemically to promote cel
Freezing－and－Thawing－cells
Freezing and Thawing cellsFreezingIt is best to freeze cells that are growing rapidly. With adherent cells, it is easiest to set up 100 mm dishes, giv
Production－of－neuronpreferential－lentiviral－vectors
实验概要Adenoviral vectors widely used to transfer foreign genes into neuronal cells possess tropism for glial cells and are toxic to infected cells.
Protocol－for－Cell－Fusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the
Culturing－Mouse－Embryonic－Fibroblasts
MaterialsTrypsin (Gibco 25200-023)3T3 Medium: 500 mL DME (Invitrogen) + 50 mL FBS (Hyclone) + 5 mL 100x Pen/Strep2x Freezing Medium: 3T3 Medium + 20%
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
The－OP9DL1－System:－Generation－of－TLymphocytes－from－Embryonic4
TROUBLESHOOTINGProblem: The OP9 cells are more than 80%-90% confluent.Solution: It is important when creating working stocks of OP9 cells for freezing
Freezing－and－Thawing－of－Mammalian－Cell－Lines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.Fre
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
Live－imaging－with－Drosophila－tissue－culture－cells1
IntroductionLive imaging provides an important complementation to the "snapshot" view obtained in fixed tissue by immunofluorescence. It allows follow
Cryopreservation－of－cell－cultures
1. Examine all flasks by inverted-phasemicroscopy. Cultures used for preservation should be grown free of antibiotics, show no signs of microbial cont
CellTrace™－CFSE－Cell－Proliferation－Kit
实验概要The CellTrace™ CFSE Cell Proliferation Kit provides a versatile and well-retained cell-tracing reagent in a convenient and easy-to-use form. The
Subculture－of－Suspension－Cell－Lines
AimIn general terms cultures derived from blood (e.g. lymphocytes) grow in suspension. Cells may grow as single cells or in clumps (e.g. EBV transform
Subculture－of－Adherent－Cell－Lines
AimAdherent cell lines will grow in vitro until they have covered the surface area available or the medium is depleted of nutrients. At this point the
Growing－Cells－in－Geltrex™－Reduced－Growth－Factor－Basement－Membrane－Matrix
实验概要Basement membranes are continuous sheets of specialized extracellular matrix that form an interface between endothelial, epithelial, muscle, or
Creation－and－Use－of－Infectious－Virus－Vector
Creation and use of your infectious vector:Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 ml of media. (This can be scaled up if desired).The f
Thawing－Cells－(Schreibers－protocol)
Thaw vial quickly in 37癈 water. Caution - vial can explode.Transfer cells to sterile, 15 mL centrifuge tube.Add 50 祃 warm FBS (fetal bovine serum, hea
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
QUALITATIVE－ANALYSIS－OF－DNA－FRAGMENTATION－BY－AGAROSE－GEL－ELECTROPHORESIS
1. IntroductionNuclear morphology changes characteristic of apoptosis appear within the cell together with a distinctive biochemical event: the endonu
CREATION－AND－USE－OF－YOUR－INFECTIOUS－VECTOR
实验概要CREATION AND USE OF YOUR INFECTIOUS VECTOR实验步骤Day 1 1. Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 mL of media. (This can be scal
TOP10－chemically－competent－cells
OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B
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
ES－Cell－Culture－and－Manipulation2
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
Dual－and－TripleCo...－（二）
主要试剂1. Blocking reagent after coating: 10% fetal calf serum (FCS) in phosphate-buffered saline (PBS), pH 7.4.2. Cells suspended in complete culture me
Method:－Lymphocyte－Transformation
Method: Lymphocyte TransformationMay 30, 1990Rosalie VeilePrinciple:Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocyte
The－OP9DL1－System:－Generation－of－TLymphocytes－from－Embryonic1
The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In VitroRoxanne Holmes and Juan Carlos Zúñiga-Pflücker1Sunn
FIXATION－and－DNA－Staining－for－Cell－Cycle－Analysis
BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to ente
RNA－FISH－on－cultured－cells－in－interphase
IntroductionFluorescence in situ hybridization (FISH) has become a widely used method in genome and molecular genetic studies. The technique is highly