GrowingOvernightCultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-capped culture tube. Large amount of culture should be made in growth flasks, using up to 25 flask volume in medium (e.g., no more than 30 mL in a 125 mL flask, or 250 mL in a 1000 mL flask).2. Touch a single colony with the end of an applicator stick and hold it in your ......阅读全文
Placental－trophoblast－and－chorionic－cell－cultures
1. Placental trophoblast and chorionic trophoblast cells were prepared using a modification of the method. 2. Term human placentae and chorion tissue
TISSUE－FIBROBLAST－CULTURES－FOR－CHROMOSOME－ANALYSIS
实验概要 Skin tissue may be used for chromosome analysis in special cases when the results from peripheral blood are inconclusive, e.g. suspect
Fungal－Genomic－DNA－Extraction
实验概要This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.High t
An－Ultrafast－method－of－DNA－extraction－from－Neurospora
We have found that the DNA extraction procedure of Metzenberg and Baitch (Neurospora Newsl. 28:20)/Stevens and Metzenberg (Neurospora Newsl. 29:27) wh
Shellless－cultures－of－chick－embryos
This experiment allows you to observe the development of a chick embryo outside its shell. Cultured chicks are also more accessible for manipulation.
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
Cosmid提取技术
Day OneStart overnight cultures of Cosmid in fresh L-Broth + 25 ug/ml kanamycin. 4 x 250 ml each, inoculate each with metal loop from frozen glycerol
Method:－Removal－of－Yeast－Contamination－from－Lymphoblast－Cultures
Purpose:This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when s
Barretts－esophageal－epithelial－and－fibroblast－primary－cultures
1. Biopsy specimens for tissue culture were immediately placed on ice in primary cell culture system. 2. Within 4 hours from the time of the biopsy, t
超级感受态细胞的制备
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Institute and T
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
Lambda（噬菌体）DNA－Miniprep
David HarryInstitute of Forest GeneticsUSDA Forest ServicePacific Southwest Research StationAugust 26, 1993Background :There are many published method
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
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
A－quick－RNA－miniprep－for－Neurospora－mycelial－cultures
Most RNA isolation techniques currently in use have been developed for the processing of large quantities of material. These typically involve multipl
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 ·
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
Primary－Culture－of－Pulmonary－Arterial－Smooth－Muscle－Cells
Primary Culture of Pulmonary Arterial Smooth Muscle Cells 1.Primary cultures of Pulmonary Arterial Smooth Muscle Cells (PASMCs) were isolated from hum
Chromosomal－DNA－Extraction－from－Grampositive－Bacteria
Chromosomal DNA Extraction from Gram-positive BacteriaThis procedure was originally developed for Listeria monocytogenes but has worked well with othe
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
Mitochondrial－DNA－Isolation－from－Somatic－Embryogenic－Cell－Cultures－of－Larix
Mitochondrial DNA is isolated by a modification of the methods described by Wilson and Chourey (1984) and Radetzky (1990). Cell cultures at four days
Primary－cultures－of－intrahepatic－bile－duct－epithelial－cells－isolated－and...
Primary cultures of intrahepatic bile duct epithelial cells isolated and cultured1. Liver was surgically removed and perfused via the hepatic vein.2.
cAMP分析－cAMP－Assays
cAMP AssaysGouzel Karimova and Daniel LadantUnite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et C
Maintenance－of－Probes－in－bacteria－including－Escherichia－coli
Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Preparation－of－Lactobacillus－Competent－Cells
OverviewInstructions on how to prepare Lactobacillus plantarum competent cells before electrotransformation.MaterialsMRS mediaCulture of L. plantarum
Human－Embryonic－Stem－(ES)－Cell－Protocols——Matrigel－Aliquoting－and－Plating
Aliquoting Matrigel:Day one:Put the sterilized tip box (either 200 ml or 1000 ml tips), sterilized eppendorf tube container, and appropriate pipettor
Rat－urinary－bladder－urothelial－cells
1. Bladders were excised from deeply anesthetized (urethane, 1.2 gm • kg−1, i.p.) Sprague Dawley rats (of either sex), cut open, and gently stretc
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
Acid－Phenol－Yeast－RNA－Prep
This is the preferred method for yeast RNA preparationuse Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fres