生物化学技术

Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small portion of glass wool at the base of the column.3) Pour gel solution into column and use a stir ...

1) Prepare a C-18 reverse phase prep-sep cartridge by washing with:a) 10 ml of methanolb) 20 ml of chloroform-methanol (2:1)c) 10 ml of methanold) 10 ml of methanol-1.6 M sodium acetate (1:1)2) Apply ...

IntroductionMilk fat measurement is a common task in the dairy industry because milk fat content is one factor that determines milk price and is necessary to know for casein/fat ratio normalization. I ...

Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.18 g50 mM sodium pyrophosphate (Na4P2O7-10H2O) 22.3 g5 mM Na2 EDTA 1.8 ...

1) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.--- If treatment is for an extended period of time: treat in serum free media containing labeled galactose.2)Wash label out and spin cells ...

Reagents:Lysis buffer25 mM Tris-HCl pH 7.45 mM EDTA1 mM ATP20 µg/ml CLAP1 mM PMSFBuffer A10 mM MgCl20.2 M Tris-HCl pH 7.40.2 % Triton X-100Buffer B0.2 M sodium acetate pH 5.00.2 % Triton X-100Buffer C ...

This procedure involves use of human plasma a potentially dangerous source of blood-borne disease. Wear long-cuffed gloves and eye-protection. Day one: -Human Plasma is purchased from the Red-Cross. I ...

As insects have an open circulatory system the hemolymph can be simply collected through an incision in the body wall. Most conveniently you should cut a leg and let the blood drip into a chilled glas ...

Synthesis reaction:1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol.2) Allow reaction to proceed at room temperature for atleast 18 hours in the dark.3) ...

NOTE - Use arachidonic acid from Biomol (# FA-003)1) Make up the appropriate concentration of AA in 100% ethanol.2) Add 1µl of phenol red solution.-- The solution should appear yellow since the free a ...

1. Make standards using sodium phosphate at the following uM concentrations: 0 2 5 7 10 20 40 60 and 80. Use the screw top glass tubes.2. Dry the samples and standards under nitrogen evaporator (Rm 20 ...

ReagentsAshing buffer:10 g Mg(NO3)2 100 ml EtOH1.5 N HCl stock:119.7 ml concentrated HCl (11.6M @ 36% by weight)880.3 ml H2O1 N Sulfuric acid stock:28.9 ml concentrated sulfuric acid (18M/36N @ 96% by ...

I. Lower chloroform phase:1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely but sufficiently to yield a final volume of ~2 mls.II. Upper aqueous phase:1) ...

Thin layer chromatography is based on the separation of a mixture of compounds as it migrates with the help of a suitable solvent through a thin layer of adsorbent material which has been applied to a ...

During this week you will analyze the fatty acid composition of the individual lipid fractions recovered from the TLC plate. Gas chromatography is a very sensitive method for the separation and quanti ...

Protocol:Bligh & Dyer extraction1) Pellet approximately 1 X 107 cells.2) Resuspend pellet in 3 ml CHCl3: CH3OH (1:2) and vortex hard.3) Add 0.8 ml H2O vortex hard and allow solution to sit at room tem ...

Lipid Extraction1) Following the appropriate time of treatment transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Spin down cells in the table top centrifuge at 1200 r ...

1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temperature PBS.3) Resuspend final pellet in an appropriate volume of prewarmed ...

Materials Tubulin (5 mg/mL) 100 mM Mg・GTP 4 mM Taxol in DMSO PM = 100 mM PIPES pH 6.82 mM EGTA1 mM Mg2SO4 Motor protein (95% purity; 15-20 µM) Cuvettes (200 µL volume 10 mm path length) 0.5 M Tris-OAc ...

Materials Nucleotide mixMotor (50 - 100 µM; purity 95%) 0.5 M Tris-OAc pH 7.5 10 mM EGTA 10 mM MgCl2 DDW Sephadex G-50 Medium column (0.8 cm in x 20 cm) Column buffer = 50 mM Tris-OAc pH 7.5 1 mM EGT ...