Ligation The objective of this experiment was to ligate EGFP DNA inserts into pET41a(+) plasmids. A total of five ligations were performed, two actual ligations and three control ligations. The following reagents were utilized: Nco I/Not I cut pET-41a(+) DNA 50 ng/μL, EGFP cDNA insert 7 ng/μL, uncut pET-41a(+) DNA/EGFP recombinant plasmid DNA 25 ng/μL, ligase buffer 10X, and ligase. To prepare ligation #1, a 1:1 molar ratio of pET41/EGFP, 2 μL digested pET41a(+) DNA, 1 μL EGFP cDNA insert, 14 μL sterile dH2O, 2 μL ligase buffer, and 1 μL DNA ligase were added to a micro centrifuge tube. To prepare ligation #2, a 1:3 molar ratio of pET41/EGFP, 3 μL digested pET41a(+) DNA, 1 μL EGFP cDNA insert, 12 μL sterile dH2O, 2 μL ligase buffer, and 1
3.9.3 Ion Exchange chromatography As maximum activity was observed in the 50-80% fraction obtained from sequential ammonium sulfate precipitation, this fraction was subjected to ion-exchange chromatography on a DEAE-Cellulose column equilibrated with 0.15 M PBS. The resin was treated and column was packed as per the manufacturer’s instructions. • 5gm dry resin was suspended in 25 ml of distilled water and left at room temperature for 30-45 minutes to settle down the matrix. The settled volume of the resin was measured and this was termed as Column volume (CV) and used as a measure for washing. • Further the filter resin was suspended in 2 CV of 0.15 M NaOH containing 0.5 M NaCl for 10 minutes, and washing was continued for 5 CV of 0.15 M
We inoculated them with our stain and incubated them on shaker for 24 hours. After 24 hours incubation, we took O.D at 600nm and plotted a graph. To check effect of different temperatures on rhizospheric nitrogen bacterial growth: We took 6 flasks (100ml), and added 20ml L-broth in each flask and autoclaved it. We inoculated them with our stain. We selected different temperatures 4°C, 15°C, 37°C.
The percentage of amplification result using the pairs of primers on the tested 56 isolates can be seen in Figure 3. The isolates which produced DNA fragment of 760 bp were 19.6 %, the isolates which produced DNA fragment of 685 bp were 30.4 %, while the isolates with the two amplicons, were 14.3 %, and 64.3 % of isolates did not have the two amplicons. The resume of phenospecies and genospecies tests using the primer 685 and the primer 760 dan be seen in Table
ABSTRACT The N-hexane, ethyl ether and ethanol extracts of leaves of Gossypium herbaceum L was investigated for anthelmintic activity using earthworms (Pheretima posthuma). Various concentrations (10, 20, 40, 60, 80 & 100 mg/ml) of plant extracts were tested in the bioassay. Albendazole (10 mg/ml) was used as reference standard drug whereas 1% v/v tween 80 as control. Determination of paralysis time and death time of the worms were recorded. The ethyl ether and ethanol extracts exhibited significant anthelmintic activity at highest concentration of 60, 80 & 100 mg/ml compared to standard drug.
It lasted 43 years until in 1993 Nadano et al. introduced a new method for DNase I activity measurement called “single radial enzyme diffusion” or SPRED (Fig.8). This method is simple and it is based on the digestion of DNA in the agarose gel by DNase, which is present in samples punched into the gel. Ethidium bromide or other currently used DNA dyes fluoresces only with
20, 40, 60, 80 and 100µl concentration of ch-agnp was added into the respective wells. . S.aureus E.coli, s.typhii, P.aeroginosa, was incubated at 37º c for 24 hrs. C.albican was incubated at 22 c at 72 hrs. Result and Disscusion: TEM Analysis- TEM microscopy illustrated that chitosan conjugated silver nanoparticles were typically in spherical shape
(Khaket et al., 2012). Vermicomposting is also a remarkable strategy for the management of Parthenium, it has also been enhance its nutrients and overcome the allelopathic capacity (Yadav and Garg, 2011). In vermicomposting, phenolic components of Parthenium is remarkably decrease, it also decrease heavy metal pecentage and toxic substances. There is significant increase in selected macronutrients(N,P,K) and decrease in organic carbon in Parthenium compost,which is suitable for organic
The total soluble protein content inside the plants can comprise up to 50%20. Because RuBisCO is so abundantly present in plants it was assumed that the same applies to microalgae. In 1991, J.A. Raven compiled a dataset for RuBisCO in microalgae as the fraction of RuBisCO to total protein by mass, showing values between 2% and 23%. To determine these percentages however, an indirect measurement was performed where RuBisCO is converted on a total protein basis assuming the chlorophyll: total protein mass ratio is 0.0421.