Method:ReactivatingCellLinesandCellGrowthforDNAPreparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extraction.Time required:15-20 minutes to begin growing 2-4 cryovialsProcedure:Frozen cells should be thawed quickly. Remove the cryovial from its long term storage container in the -135 degrees C Cryostar, and place immediately in a 37 degrees C water bath for 2 minutes.Rem......阅读全文
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Method:－Maintaining－Lymphoblastoid－Cell－Lines
Method: Maintaining Lymphoblastoid Cell LinesJune 10, 1990Rosalie VeilePurpose:To grow lymphoblastoid cells for permanent storage and for DNA extracti
细胞培养——细胞保藏
Working Cell Bank (Contributed by Nanci Donacki)Provides detailed protocol for establishing a working cell bank Master Cell Bank (Contributed by Na
Method:－Preparation－of－Lymphocyte－Cell－Pellet－for－Storage
Method: Preparation of Lymphocyte Cell Pellet for StorageJune 10, 1990Rosalie VeilePurpose:Following propagation to 1 X 108 cells, lymphoblastoid cell
Method:－Lymphoblastoid－Cell－Lines－from－Frozen－Whole－Blood
Method: Lymphoblastoid Cell Lines from Frozen Whole BloodMay 31, 1990Rosalie VeilePurpose:Blood Samples can be stored frozen as a backup in case an LC
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
A－practical－method－for－the－preparation－of－total－DNA－from－filamentous－fungi
Most methods of DNA preparation from fungi are time-consuming due to the need to first make protoplasts, expensive for chemicals such as cesium chlori
Easy－YAC－Preparation－Method
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw
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 ·
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man
Cryopreservation－of－Cell－Lines
AimThe protocol below describes the use of passive methods involving an electric -80ºC freezer for the cryopreservation of cell cultures. ECACC routin
Resuscitation－of－Frozen－Cell－Lines
AimMany cultures obtained from a culture collection, such as ECACC, will arrive frozen and in order to use them the cells must be thawed and put into
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
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
RNA提取
RNA提取（主要内容如下）Tips for Handing RNA Total RNA IsolationmRNA IsolationrRNA IsolationOthersQ & A posted in the Method Forum Basic Procedures for Handing
Method:－Logging－in－Specimens－and－Record－Keeping
Method: Logging in Specimens and Record KeepingJune 10, 1990Rosalie VeilePurpose:To keep a written and computerized record of all cell lines, the date
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
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells1
Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the c
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells2
Flow Cytometric AnalysisPrior to flow cytometric analysis, the vital dye Hoechst 33342 (Sigma-Aldrich, St. Louis, MO, USA) was added to cell suspensio
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells3
Moreover, when the method was applied to the analysis of the cellular composition of immature testes, the results were also in agreement with previous
Competent－Cell－Preparation
实验概要Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. These cells readily incorporate f
CELL－MEMBRANE－PREPARATION
I. Solutions: A. Ca and Mg free Phosphate Buffered Saline (PBS) solution, buffered with 0.02M Hepes. pH=7.4 B. Ca and Mg free PBS, buffered with
Subculture－of－SemiAdherent－Cell－Lines
AimSome cultures grow as a mixed population (e.g. B95-8 - marmoset) where a proportion of cells do not attach to the tissue culture flask and remain i
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Generating－stable－cell－lines－in－HEK293
Generating stable cell lines in HEK293Prior to transfection, it is recommended that you linearize your pcDNA gene construct. Linearizing will decrease
Method:－Cell－Counts－Using－a－Hemacytometer
Method: Cell Counts Using a HemacytometerJune 1, 1990Rosalie VeilePurpose:The purpose of this procedure is to determine the cell density of the cultur
巨噬细胞和单核白细胞
· Lymphocyte Transformation (Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
DNA转化
DNA转化Chemical Transformation· Transformation of Competent Cells (RbCl2 Method) (Goldberg Lab)Very nice protocol for E. Coli transformation inc