After the finalization of alkali formulation to be used for extraction of cornhusk fibres, the fibres were treated with Pulpzyme HC to increase their fineness. Pulpzyme HC is a xylanase enzyme (EC.184.108.40.206) produced by submerged fermentation of a genetically modified Bacillus microorganism, and was obtained from Novozymes. The enzyme has an activity of 1000 AXU per gram. (http://fzfz.nbdl.gov.cn:81/files/20130815/1376527490502_36.pdf) Effect of concentration of Pulpzyme HC and Treatment Time The extracted fibres were treated with three different concentrations of enzyme i.e. 1%, 3%, 5% (owf).The time taken for treatment was 30 minutes, 60 minutes and 90 minutes.
MATERIALS AND METHODS Bacterial strains and culture conditions Two S. aureus strains were used in the present study; S. aureus 8325-4 (SigB-) and SH1000 representing a SigB+.strain. Overnight cultures were grown in Luria Broth (LB) at 37°Cwith shaking at 150 rpm. Exposure to antibiotics was carried out as detailed below. Antibiotics Ciprofloxacin were purchased from Sigma-Aldrich CO. 10 mg/ml stock solution of antibiotic were prepared freshly with 0.1N HCl and stored at -20°C. During the experiment we diluted with sterile water 1:10 and 1:100 depending on the different drug concentration.
21 Fresh semen samples were diluted with phosphate-buffered saline (PBS) to 2x106 sperm/mL. Fifty µL were incubated with 100 µL lysing reagent for 15 seconds and then 2 mL of PI was added and mixed with tube. Tube acquisition was done by flowcytometry immediately after staining. The intensity of its emission corresponds to the DNA content. Flowcytometric analysis displays a constant and characteristic bimodal nonartifactual DNA pattern indicating the presence of two different populations.
22.5 g of plate count agar powder was dissolved in a litre of sterile distilled water on the hot plate 2. pH of the solution was adjusted to 7.0 ± 0.2 by adding NaOH or HCl and was immediately transferred into the Schott bottle to be autoclaved at 121 ° C for 15 minutes 3. Prepared medium was stored in 4° C chiller Lauryl Sulphate Broth
The dried mushroom samples were further heated at 105 ºC overnight until constant weight obtained for moisture content determination. The samples were incinerated at 550 ºC for 24 h and reweighed to determine ash content. MicroKjeldahl method was employed for the crude protein content (N × 4.38). The crude fat was determined by extracting a known weight of sample in Soxhlet apparatus using petroleum ether as a solvent. Total carbohydrates calculated by the difference.
H and S were diluted 20x and P1 and P2 were diluted 2x to make up a total volume of 1ml each. 50ul of each diluted sample was pipetted into 8 wells of the microplate and 50ul of each protein standard was pipetted into 2 wells. 50ul were incubated with 50ul of alkaline copper reagent. 50ul of alkaline copper reagent was added to every well containing water, buffer, sample or standard and was incubated for 30 minutes. 100ul of Folin reagent was added to the wells and incubated for another 20 minutes.
TCBS agar was prepared for about 100ml and poured into petri plates, after solidifying, pure colonies of bioluminescent bacteria was streaked. The petri plates were observed after 24 hours. If there is presence of yellow color colonies, it is concluded as Vibrio spp. 3.12.4 Species Verification by Sub-culturing at 4°C using SWC agar media The pure culture of Bioluminescent bacteria was streaked on the agar media and refrigerate at 4°C. The Photobacterium phosphoreum will exhibit slow growth at
The DEAE CL-6b gel was washed twice with 0.5m Hcl, twice with 0.5m Naoh and twice with PB ph6.0 before utilization. For DEAE 1ml gel every 1ml serum was utilized. In the wake of blending for 1 hour at 200c the suspension was centrifuged at 4500g for 25 minutes. The supernatant was thought by Ammonium sulfate precipitation and pellet was broken up in refined water and the ensuing arrangement was dialyzed against PBS ph7.2 to evacuate Ammonium sulfate and conformed to the first volume with yhe same cushion (Hong et.al., 1994). 2)
H2SO4 • Catalyst: potassium sulphate (K2SO4), copper (II) sulphate pentahydrate (CuSO4.5H2O) • NaOH (40%) • 0.1 N HCl solution • Boric acid (4%) • Indicator: methyl red (200 mg make up to 100 ml with %95 ethanol ) 1 gram Feta Cheese was used as a sample in this experiment. First of all, the experiment was started with the digestion phase which was performed by mixing 10g potassium sulphate and 0.5 g CuSO4 solution that acts as catalyst, and added 20ml conc. H2SO4. The digestion unit temperature was set to 420±10°C and preheated for 15 minutes. Then, it was heated continuously at least 2 hours till a clear colorless solution was obtained.
We added 50ml of the buffer into the flask, and we boiled the solution for two minutes in the microwave. The sample well was taped with table and par film to keep the agarose gel from leaking. I loaded the 15ul of the five serum proteins: Cow serum, Serum albumin, Transferrin and Gamma globulins. The electrophoresed gel was placed on the voltage machine at 110 volts for 1.5hours. We removed the gel from the sample well and place the Coomassie Blue stain.