PouringPlates
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 than the media volume.2. For minimal medium, make separate 2X agar and 2X salt flasks. For example, for 1 liter of E plates, make 500 mLs of 2X E salts, and 500 mLs of 2 X agar (1.5 g per 500 mLs). Use at least one flask large enough to mix the media together after autoc......阅读全文
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
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
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
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
Marcantonio－Lab－Protocol－Manual
Protein GelPreparing and Running a Protein Gel (7% Polyacrylamide) A. Preparation of Running Gel Solution 1) Add to a 50 ml cylinder:DD-H2O 26 ml30% a
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
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
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
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
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
DualColor－ELISPOT－Assay－for－the－Simultaneous－Detection2
21. Add 100 µL of developing buffer to all wells using a multichannel pipette and incubate at room temperature for at least 2 h. 22. Wash the ELISPOT
SSR－GEL－and－Silver－Staining－Protocol
I. EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
Sterile－Technique
Good sterile technique is the first and most important step in insuring consistent results when employing recombinant DNA and protein expression techn
CHO－Centrosome－Prep
CHO Centrosome Prep:Arshad Desai4/94Cells:We grow our CHOs with MEM[[alpha]] (without nucleosides) + 10% Bovine Calf Serum and penn/strep/glutamine. F
无菌化技术
Sterile TechniqueGood sterile technique is the first and most important step in insuring consistent results when employing recombinant DNA and protein
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
In－vitroculture－of－early－chick－embryos
1. Use sterile technique. Prewarm Howard's Ringers in petri dish andagar/albumin culture dish to 37ºC.2. Crack 2-day egg into large sterile petri
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
DNA－Fragmentation－Assays－for－Apoptosis
Protocol I: Triton X-100 Lysis BufferIn 96 flat-wells plate, incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of eff
DNA测序
DNA测序（主要内容如下）· Sequencing Gel Preparation· Preparation of Templates · DNA Sequencing by the Dideoxy Method· DNA Sequen
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: