Basicproceduresforbacteriaculture1

A. Phenol extraction of DNA samplesPhenol extraction is a common technique used to purify a DNA sample (1). Typically, an equal volume of TE-saturated phenol is added to an aqueous DNA sample in a microcentrifuge tube. The mixture is vigorously vortexed, and then centrifuged to enact phase separation. The upper, aqueous layer carefully is removed to a new tube, avoiding the phenol interface and then is subjected to t......阅读全文

Fluorescence-Procedures-forthe-ActinandTubulin-Cytoskeleton-inFixed-Cells1

General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,

Fluorescence-Procedures-forthe-ActinandTubulin-Cytoskeleton-inFixed-Cells2

Actin CytoskeletonMethanol fixationFix in -20oC methanol for 1-2.5 minutesRinse in TBSPermeabilize in TBS-0.5% TX for 10 minutesRinse in TBS-0.1% TX (

麦康凯琼脂平板

This image illustrates growth of a nonlactose fermenter on MacConkey agar. MacConkey agar contains bile salts and crystal violet which inhibit the

Cell-Sorting-By-FACS

Currently, the Moflo instrument (Dakocytomation) is used to sort cells at the AECOM FACS facility.Sorting is performed by the person in charge at the

MS-Plant-Tissue-Culture-Medium

Component mg/l in MS mg/l in stock Amount for

Isolation-and-culture-of-smooth-muscle-cell

1.Male Wistar rats of about 250g body wt were anesthetized with ether and were decapitated.  2.Kidneys and thoracic aorta were removed, rinsed with

Formaldehyde-Treatment-of-Tissue-Culture-Hoods

You will need:-12g Potassium Permanganate 6g Crushed paraformaldehyde1) Set the hood so that it can vent outside.2) Mix the two chemicals above togeth

TISSUE-CULTURE-STOCK-SOLUTIONS-AND-MEDIA

MS MEDIUM FOR ARABIDOPSISTo 990 ml H2O add: Sucrose ........... 10.0  g MOPS ..............  0.5  g Agar ..............  8.0  g Adjust pH to 5.7

FeederFree-Culture-of-hESCs

MEF Conditioned mediumPlate MEFs (p4 or p5) onto gelatin-coated plates at a density of 1x106 cells/10cm culture dish.The next day, wash cells with PBS

Culture-of-Endometrial-Explants-to-Monitor-Synthesis

Reprinted from:Hansen, P.J and Betts, J.G. (1992) Culture of endometrial explants and peri-implantation conceptuses to monitor synthesis and secretion

Tissue-Culture-Methods1

I. TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often us

Isolation,-Culture,-and-Differentiation-of-Progenitor-Cells

Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous SystemScott R. Hutton and Larysa H. Pevny1UNC Neuroscience Center

Tissue-Culture-Methods2

IV. MAINTENANCECultures should be examined daily, observing the morphology, the color of the medium and the density of the cells. A tissue culture log

Human-B-cell-isolation-and-culture

B cell isolation1. Donor blood was obtained with informed consent under guidelines issued by the Medical Ethics Committee. 2. Mononuclear cell fractio

Exercise-12.10--Establishment-of-a-Primary-Culture

Exercise 12.10 - Establishment of a Primary CultureLEVEL IIIMaterialsChick embryo (approximately 8 days old)70% (v/v) ethanol for swabbingSterile scis

Follicular-thyroid-cell-isolation-and-culture

Thyroid epithelial cells (also called follicular cells or principal cells) are cells of thyroid gland that are responsible for the production and

Human-B-cell-isolation-and-culture

实验概要This protocol provides a general protocol for human B cell isolation and culture.实验步骤B cell isolation1. Donor blood was obtained with informed con

Primary-brain-cell-isolation-and-culture

1. Cerebella were removed from 7-day-old mice and passed through Nitex nylon netting (80 μm pore size) into primary cell system containing 20% (v/

Isolation-and-culture-of-pancreatic-stellate-cells

Pancreatic stellate cells (PaSCs or PSCs) are myofibroblast-like cells found in the areas of the pancreas that have exocrine function.  PaSCs are

Immunofluorescence-Microscopy-of-tissue-culture-cells

Immunofluorescence Microscopy of tissue culture cellsThese methods are written for direct staining of filamentous actin with bodipy FL-phallicidin and

Culture-of-retinal-endothelial-cells-and-pericytes

Isolation of retinal microvessels1. Eyes were obtained and transported on ice.2. Eyes were cut circumferentially 3 mm posterior to the limbus, the v

Isolation-and-culture-of-porcine-muller-cells

1. Porcine eyes were harvested from pentobarbital-anesthetized animals, each weighing 30 to 40 pounds, and transported to the laboratory in ice-co

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

Lactobacillus-transformation

OverviewThis page details a electrotransformation protocol for Lactobacillus bacteria, specifically Lactobacillus delbruckii subsp. bulgaricus and Lac

Transfer-of-Eukaryote-Suspension-Cultures

MaterialsFibroblast suspension cultureTissue culture laminar flow hoodMedia appropriate to culture line usedDisposable pipettes (10 ml and 1.0 ml)Disp

Preparation-of-Plasmid-DNA-by-Alkaline-Lysis-with-SDS:-Maxipreparation

实验概要Plasmid DNA is isolated from large-scale (500 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and SolutionsAlkaline lysis solu

细胞培养——培养基和试剂

·         Cell Culture Media and Solutions (Donis-Keller lab)·         Cell Culture Media Formulations (LTI)Comprehensive list of cell culture media,

Isolation-of-human-multipotent-mesenchymal-stem-cells-from-second

Isolation of human multipotent mesenchymal stem cells from second‐trimester amniotic fluid Culture of MSC from amniotic fluid1. Twenty amniotic flui

Bacterial-Colony-PCR

Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr

General-Cloning-Protocols

Large Scale Preps: (See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a.m. w