the supernatant was discarded and the pellet re-suspended in 400 µl of TE buffer (40 mM Tris-Base, 20 mM acetic acid, 1 mM EDTA, pH 8.0). 6 µl of 7.5 M ammonium acetate was added and the pervious step was repeated. 700 µl of 70 % ethanol was added and spun for 2 min at maximum speed. The supernatant was discarded and dried at 37°C, then finally DNA precipitate was re-suspended in 100 µl of TE
After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes. These were centrifuged at 3000 rpm for 15 minutes. The absorbance of the supernatant was read at 540 nm at room temperature against appropriate blank. Blank consist of 1 ml distilled water, 0.5 ml of 30% TCA, and 0.5 ml of 0.8% TBA. TBARS values were expressed as n moles malonaldehyde (MDA)/mg protein.
This organism was cultured under solid-state fermentation for 72 h using wheat bran as the substrate. After 72 h, crude enzyme was extracted from the culture medium. The fibrinolytic enzyme was purified from the crude sample by various steps: ammonium sulfate precipitation, dialysis, ion exchange chromatography, and casein-agarose affinity chromatography. All purification steps were performed at 4°C until otherwise stated. The crude enzyme was precipitated at 70% saturation of ammonium sulfate, and the protein was collected by centrifugation (10,000×g for 10 min).
The equipment used for preparation the extraction unit are shown in Figure 1. The pH of aqueous solution containing 10 μg L−1 of Cd (II) and 50 μg L−1 of Pb (II) was adjusted with HNO3 and NaOH (0.1 M). Then 2 mL of this solution was poured into the syringe. Afterward, 2 mg of NDNPG was added to the extraction unit and immersed in an ultrasonic bath for 60 s to disperse the NDNPG in the sample solution and increase the contact surface between the adsorbent and solution. After sonication, NDNPG was separated from sample solution easily by pressing the plunger, the sample solution from the syringe came out and NDNPG adsorbent remain in the syringe.
Plant material and extraction 300gm dried powder of seed was weighed & taken in a aspirator (2.5L). Before placing powders into the aspirator, the jar was washed properly with acetone and then dried. 800ml of solvent i.e. methanol was added gradually. The container with its content was sealed & kept for 20 days with occasional shaking & stirring.
After the evaluation of stomach for ulcers, the gastric mucosa of glandular portion was scrapped with the help of two glass slides, weighed (100 mg) and homogenized in 1 mL of a 0.15 M, ice cold potassium chloride (KCl) solution and centrifuged at 3,000 RPM for 10 minutes (REMI centrifuge). 1 mL of suspension was taken from the above tissue homogenate in test tube and 0.5 mL of 30% w/v TCA (trichloroacetic acid) was added to it, followed by 0.5 mL of 0.8% w/v TBA (thiobarbituric acid) reagent. The tubes were then covered with aluminium foil and kept in water bath for 30 minutes at 80 °C. After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes and centrifuged at 3000 rpm for 15 minutes (R-BC DX REMI centrifuge). The absorbance of the supernatant was read in spectrophotometer (UV-1601, SHIMADZU) at 540 nm against blank.
The column was centrifuged empty at full speed for 1 min to remove any residual ethanol. The RNA was eluted by adding nuclease-free water into the column and centrifugation of 1 min at full speed. The yielded RNA was checked by nanodrop (Thermo) and the integrity of the RNA was assessed in the Experion automated electrophoresis station
Kashibai Navale college Of Pharmacy, Kondhwa Bk. Preparation of Leave Extract 50 g of Psidium guajava linn leaves are washed with distilled water and keep in dark room for 4 days and grind into powder form. 2 g of powder dissolved in ethanol, methanol, pet ether of 50 ml of solvent respectively in incubator at 40oc for 5 days. The extract was subsequently filtered and concentrated to dryness. Antibacterial assay The screening was done by disc diffusion method.
(2011), who reported statistically significant increase in plasma activities of ALT, AST, ALP, and GGT in CCl4 induced animals. It was observed in this study that the feeding of rats with peppermint and parsley leaves oils was able to reduce and sometimes completely remove the toxic effect of CCl4. Some parameters including ALT and AST were significantly decreased (p< 0.05) in the group injected and fed with CCl4 and peppermint or parsley leaves oils compared with CCl4 group. Frank et al., (2012) reported that CCl4 produces oxidative damages and increased AST, ALT, ALP, total bilirubin levels and decrease total protein. Therefore, the reduction in serum levels of AST, ALT, ALP by treatment peppermint and parsley leaves oils is an indication of stabilization of plasma membrane as well as repair of hepatic tissue damage caused by CCl4.
Not enough time was given for the precipitate to settle in the beaker, hence when the supernatant is removed; there is a large amount of product lost. This is especially so in step 25, because a large amount of solvent is decanted. It will require much longer than the given time of 10 minutes to allow most of the precipitate to settle at the base of the beaker. Hence instead of decanting the mixture, centrifugation can be used to remove the solution instead. The amount of product lost will be significantly lower by carrying out this process.