常用设备/其他

Each time the microscope is to be used it should be set up correctly to give a good image. Most often users forget to adjust the iris diaphragm and focus the condenser to give its optimum performance, this could result in objects in the preparation being undetected. There are many microscope set up procedu ...

Materials Needed20ml glass scintillation vial Small stir bar Foil Glycerol 1X PBS Pipets * P-phenylenediamine ( EMD Chemicals Inc. Cat# PX0730) Carbonate-Bicarbonate Buffer (see below) Wrap a glass scintillation vial with foil and drop in a small stir bar. (PPD is light-sensitive.) With a 10 ml p ...

Good sterile technique is the first and most important step in insuring consistent results when employing recombinant DNA and protein expression techniques. Sterile technique refers to procedures by which cultures may be manipulated without infecting the worker or contaminati ...

Hahn LabCheck manuals for more detailed information.Please Note: We have been having problems with the seal and bearing assembly at the bottom of the unit which allows the shaft to rotate inside the growth chamber. It is important to always have seal steam pressure (10-15 psi) when stirring the media, ...

PEAS Labeling Gcn4-cysJames Fishburn, Hahn Lab March 2006Notes10 mCi 125I = 4.6 nmol (see spec. sheet for calculation)4.6 nmol Gcn4(1-118,209-281) = 103 microgram (M.W. 22,400)PEAS has low solubility in aqueous solutionMinimize the time Gcn4 is without DTT by preparing the protein just befo ...

Ampicillin (50mg/ml)Ampicillin Dissolve Ampicillin (Na salt from Sigma, A-9518) in H20. Store 1 ml aliquots at 20oC in common freezer for stock solutions 上一篇：PREPARATION OF PROTEIN A SEPHAROSE CL 4B BEADS 下一篇：ALPHA FACTOR ARREST AND RELEASE

Blotting with BSA Blot in 5% mile/ 1X PBST (or 1X TBST) --60'/Rt Wash 2X for 5' in 1XPBST 10Ab in 1% BSA/1X PBST --60'/RtWash PBST 15', then 3x for 5' each in 1X PBST20Ab in 1% BSA/1X PBST --60'/Rt (aHRP)--Wash 15' with 1X PBST then 3x with PBST for 5' @Chemiluminescence: Mix 1:1 NEN reagents. Add to Mb --1'/shaking. Blot excess Put between s ...

GLASS BEADS0.45 - 0.5mm beads (Sigma)Clean by soaking in nitric acid --stir bar, O/N.****Note, dispose of acid in appropriate flask.Wash w/ lots distilled water (til pH to that of water, allow to soak ~2H between changes to ensure clean)Dry before use (bake in oven)Microbeads: Cataphote Inc. 1-800-221- ...

Oxalyticase Dissolve in 0.1 M NaCl, 0.02% Na azide, 50% glcyerole Store aliquots (1 ml) at 20oC in common freezer for stock solutions 上一篇：ALPHA FACTOR ARREST AND RELEASE 下一篇：GLASS BEADS

Alpha Factor Arrest and ReleaseAlpha factor stock is 10-3 M in 0.1N HCl (i.e. resuspend 5 mg alpha factor in 2.97 ml 0.1N HCl). To arrest BAR1 strains, the pheromone should be diluted to a final concentration of approximately 3 x 10�5 M, while bar1 strains require much lower pheromone concentrations (10-8M). To ...

RNAase Dissolve in 50 mM KAC pH 5.5 Heat to 100 oC for 15 minutes and cool slowly to room temperature Store at -20. (common freezer for stock solutions) 上一篇：GLASS BEADS 下一篇：Salmon Sperm DNA (10mg/ml)

SCE 0.5 M Sorbital 0.05 M Na Citrate 0.005 M EDTA pH to 5.8 上一篇：Salmon Sperm DNA (10mg/ml) 下一篇：SPECTROPHOTOMETER INSTRUCTIONS

2nd Floor Spectrophotometer w/ monitorTurn on power switch. Bring up Menu (hit Mode button?) at top of screen; it asks if you want to make changes? Y or N. Hit Yes. Hit the number 1. (for changing wavelength) Hit Enter. type in wavelength ex. 260 Hit Enter. Now you're back at the main Menu which asks if you want to make changes ? Y or ...

POSI BLOT SOLUTIONS2X Posi blot denaturing sol'n2 Liters80g NaOH sodium hydroxide (pellets)350.6g NaCl sodium chloridebring up to 2 liters with H2Ostir until dissolvedaliqot into two 1-liter bottlesdo not autoclave2X Posi blot neutralization sol'n2 Liters484.4g Tris basebr ...

Zymolyase Dissolve in 10% glucose. Store aliquots (25 µl) at 20oC in common freezer for stock solutions Use once and dispose Comments: alternative is to dissolve in SCE, 40% gycerol, 20 mg/ml final. Can use multiple times. 上一篇：POSI BLOT SOLUTIONS 下一篇：10X TBE BUFFER

10X TBE BUFFER3 Liters324 g Tris base165g Boric acid120mls 0.5M EDTA (pH 8.0)autoclave for 20 min2 Liters216g Tris base110g Boric acid80mls 0.5M EDTA (pH 8.0)autoclave for 20 min1 Liters108g Tris base55g Boric acid40mls 0.5M EDTA (pH 8.0)autoclave for 20 min 上一篇：Zymolyase (3mg/ml) 下一篇：20X SS ...

20X SSPE BUFFER3 Liters525.9g NaCl sodium chloride82.8g NaH2PO4 sodium phosphate monobasic28.2g EDTA powder FW=372bring up to 2400mls with H2Oadd NaOH to pH 7.4 (~27mls/liter of 10N NaOH)autoclave for 20 min 2 Liters350.6g NaCl sodium chloride55.2g NaH2PO4 sodium phosphate monobas ...

0.5M EDTAFWMLpH8.0xg =292.250.50.undefined*93.06undefinedAdd_to_about_300mls_H2O_while_spinningph_to_8.0bring_up_to_500mlsfilter_sterilizenote~I_FW~I_372.24~J_M~I0.5~J_L~I_0.5~J_**93.06undefined_________上一篇：20X SSPE BUFFER 下一篇：CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEAST

ChIP Protocol-Mechanical Breakage & FA Lysis BufferDay -2:o Start a 5ml overnight culture from a single colony.Day -1:o Start a larger overnight culture using part of the 5ml culture (subculture).Day One - Part One:1. Grow culture until it is in log phase: OD600=0.7 - 1.22. For each sample you collect, put t ...

Flow cytometry of yeast cells stained with PIShort Coursefill flow buffer tank, empty waste turn on FACSCalibu turn on computer double click on "Bob's FL3h" file on desktop (an error message will come up - hit "OK") under the Aquire menu: designate the file folder location and file name in Parameter Desc ...