Layeredplates
General InformationThe cells being plated are sandwiched between layers of cell-free agar. The submerged cells grow into smaller colonies and many more colonies can be counted per plate (I''ve been able to distinguish over 1000 yeast colonies per plate using this method--Jgritton 14:46, 22 Aug 2005 (EDT)). In addition by avoiding the use of a spreader you can eliminate the error due to cells sticking to ......阅读全文
Layered－plates
General InformationThe cells being plated are sandwiched between layers of cell-free agar. The submerged cells grow into smaller colonies and many mor
Pouring－Plates
1. For rich media, weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger t
Dropout－plates－for－yeast
Materials(Solutions are all available from the media room)200ml bottle of 2x SD200ml bottle of 4% agar -- make sure to sign it out40% glucoseCSM minus
Preparation－of－Agar－plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow the s
Agar－Plates－for－Selection－of－Clones－in－Bacteria
Cloning of PCR productsStocks:LB Agar: Luria Broth after Lennox:per LiterTryptone10 gYeast Extract5 gSodium Chloride5 gBact. Agar15 g pH 7.0Autoclave
Preparation－of－Broth－and－Plates,－etc.
Recipes: 1) LB BrothMake 16 gm of LB Broth Base (Gibco #M27800C) up to 800 ml in ddH2O. Swirl to dissolve, then add 110 µl of 10 N NaOH. Autoclave. 2
Preparing－Antibiotics－Stock－Solution－and－Ampicillin－Agar－Plates
AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore comme
A－protocol－for－cleaning－and－reusing－the－large－25－x－25－cm－plates
We regularly reuse our large 25x25 cm plating trays; initially, however, we were plagued by gross microbiological contamination when reusing the trays
Exposing－gels－and－plates－containing－radioactive－samples－to－Xray－film
Although most people use the PhosphorImager for western blots, kinase assays and methionine-labeled samples, X-ray film remains the best way to expose
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.
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
Maintenance－of－Cell－Culture
Maintenance of Cell CultureAuthor: Nanci DonackiSource: Contributed by Nanci DonackiDate Added: Tue May 14 2002Date Modified: Tue Apr 27 2004Abstract:
RNAi在细胞培养中的应用
The protocols listed here are for Drosophila cells in 6 well plates and our pre-aliquoted 384 well plates. RNAi experiments may be done in other size
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
Preparation－of－Polyacrylamide－Gels
1. Prepare 20X TBE as:216 g Tris Base110 g Boric Acid80 mL 500 mM EDTA, pH 8.0700 mL ddH2OMix. Bring volume to 1 L. Autoclave.2. Prepare Acrylamide so
Detection－of－Mycoplasma－by－Culture
AimDetection of mycoplasma by culture is the reference method of detection and has a theoretical level of detection of 1 colony-forming unit (cfu). Ho
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g
新技术：InCell－Western－Assay2
III. Experimental ConsiderationsProper selection of microplates can significantly affect the results of your analysis, as each plate has its own chara
Cloning－by－Limiting－Dilution－of－Hybridoma
Author: Nanci DonackiSource: Contributed by Nanci DonackiDate Added: Tue May 14 2002Date Modified: Tue Apr 27 2004MaterialsDMEM, high glucose (Life Te
384well－PCR实验方法
Beforehand.choose empirically the annealing temperature of primers;make sure you have enough PCR-plates, Q-covers and sealing tape;prepare the stocks:
Marcantonio－Lab－Protocol－Manual——3
Sequencing GelPreparing and Running a Sequencing Gel (6% Polyacrylamide/Urea) A. Preparation of Gel Solution 1) Weigh out 50 g of Urea into a clean 25
Dual－ELISPOT
实验概要We provide a protocol using an FITC-conjugated primary antibody and a biotinylated primary antibody. Those are in turn recognized by anti-FITC H
Transformation－of－Electrocompetent－E.－coli－with－Blue/White－selection
Desalt DNA template by EtOH precipitation in NaOAc followed by at least 2x washes with 70% EtOH. Resuspend in 5 - 15 µL of sterile H2O.Rinse cuvettes
Fusion－and－Cloning
ReagentsMedium A - Pre-fusion Medium and Hybridoma Expansion MediumMedium B - Fusion Medium Medium C - Hybridoma Recovery MediumMedium D - Hybridoma S
Fusion－and－Cloning
Author: Nanci DonackiSource: Contributed by Nanci DonackiAbstract: Procedure for establishing hybridoma in one stepReagents(StemCell Technologies, Inc
Twohybrid－analysis－of－genetic－regulatory－networks
1. Introduction and BackgroundThere is a great need for general methods to characterize the proteins that contemporary biology makes available. The li
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble
Yale－Immunofluorescence－Protocol
实验概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要试剂Reagents1. 384-well view plates (Aurora)2. HUVEC (pooled, L
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and OverlayDescription Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str