2.5. Bovine Serum Albumin (BSA) binding Studies: BSA solution was prepared in 5 mM Tris-HCl/10 mM NaCl buffer solution to keep pH value constant (pH=7.4) and it is stored at 4oC, used within 2h. The BSA concentration was estimated spectrophotometrically (ε280=44,300 M-1cm-1) [35], 60 µM BSA was titrated with increasing concentrations of Ru (II) complex (0-50 µM). Absorbance changes at 280 nm were monitored by UV-spectrometry. 2.6. DNA Binding Studies: Binding studies of DNA were carried out at room temperature. The tris buffer (5 mM Tris-HCl, 50 mM NaCl, pH=7.1) was prepared using doubly distilled water. At fixed concentration of Ru(II) complexes(10 μM) in the buffer, by increasing the DNA concentration absorption spectra were recorded. The binding strength of the complex …show more content…
[DNA]/(Ԑa-Ԑf) versus [DNA] graph was plotted. Slope of the straight line gives Kb …show more content…
All the complexes were tested for their antimicrobial activity against viz. E.coli, Bacillus subtilis and Staphylococcus aureus. Two different concentrations, 1mg/ml (1000 μM) and 0.5 mg/ml (500 μM) in DMSO were used for testing spore germination of each fungus. 5 mm size filter paper disks were prepared using whatmann filter paper no.1 saturated with 10 μl of complexes and placed in petri dishes containing LB (Luria Bertini) agar media inoculated with E. coli, Bacillus subtilis and Staphylococcus aureus separately. The petri dishes were incubated at 25±0.2°C and the zone of inhibitions were noticed after 48h. The plates were observed and the diameters of the inhibition zones (in mm) were deliberated, and compared with standard antibacterial drug
Experiment VIII was performed to analyze SN2 and SN1 using tertiary and primary substrates and use gas chromatography (GC) to examine the SN1 reaction. The product of the SN2 reaction was classified as n-butyl iodide by using infrared spectroscopy and gas chromatography mass spectroscopy and the product of the SN1 reaction was identified as of t-butyl chloride by using infrared spectroscopy and gas chromatography. For the SN2 reaction, 7.62 grams of n-butyl bromide, 20.0 grams of sodium iodide, and 79.1 grams of acetone were used to produce 3.12 grams of n-butyl iodide. The limited reagent was identified as n-butyl bromide and the theoretical yield of n-butyl iodide was calculated as 10.3 grams. The percent yield of this reaction was calculated
The goal of this experiment is to see the anti-cow antibody bind to cow serum only, and we expect to see the anti-cow antibody bind to the spot that had the cow serum. The system we used is the serum from Cow, Horse, Goat, Sheep, and Donkey, Chicken. In order to able to detect and analyze proteins based on their ability to bind to a specific antibody, the SDS-PAGE and Western Blot was performed. SDS Polyacrylamide Gel Electrophoresis (SDS-PAGE) is a very common technique used to separate proteins by molecular weight under the influence of an applied electrical field and then used to prepare for the Western Blot (#1 Lehninger). It uses a polyacrylamide gel as a support medium and sodium dodecyl sulfate (SDS), which is a detergent, to denature
They all lack the enzyme to help determine the absorption of just the enzyme.
One of the major concerns for people, is the growing amount of wrinkles on the skin. While one can try to lose weight, and look fit, it is not easy to fight with the aging process. In fact, women, who are 25 years or older, start facing the ill effects on their skin, due to dehydration. But, with so much of advancement in the field of medicine, nothing is impossible anymore. And, there are easy ways to get rid of those ugly wrinkles, without getting under the knife.
The Kirby-Bauer disk diffusion method is utilized as a part of this trial so as to watch the viability of a sure anti-microbial. This strategy incorporates the spreading of the two sorts of bacteria being concentrated on an agar in a petri dish with the goal that they can produce. One agar contained Wild S. Aureus and another agar contained MRSA. Paper circles are let to absorb refined water, and equivalent centralizations of the accompanying antibiotic arrangements: Methicillin, Vancomycin and XR21347. At that point, these paper plates are put onto both agars and left as a base for bacterial development.
Inducing Prodigiosin Transposon mutagenesis in Serratia Marcescens Introduction Serratia Marcescens is an opportunistic pathogen, mainly of healthcare facilities but can also be found in many diverse environments. Serratia is a gram negative bacteria which can give it innate resistance to certain antibiotics, especially those that target peptidoglycan cell wall synthesis, due to its outer membrane. In an environment with different microorganisms competing for food Serratia holds a component that gives it another selective advantage. The bacteria contains a red pigment called prodigiosin, that has antibacterial, antifungal, and even antiprotozoal activity.
Tn 4351 was originally isolated from bacteroides fragilis [30] . The transposon was successfully introduced into Cytophaga succinicans, Flavobacterium meningosepticum, Flexibacter canadiansis, Flexibacter strain SFI and Sporocytophaga myxococcoides by conjugation [25]. Tn 4351carries two antibiotic resistance gene. One of the codes for resistance to erythromycin and clindamycin which is expressed in bactroides but not in E.Coli. The other gene codes for resistance in tetracycline and is expressed in aerobically grpwn E. coli, but not in anaerobically grpwn E. coli or in bacteroides.
In this three-week long experiment conducted in the Bio 13 Lab, we were able to analyze a single nucleotide polymorphism (SNP) in our own genomic DNA and then determine our genotype at this specific SNP. In week one, we extracted genomic DNA from our cheek cells with swabs and prepared our DNA for PCR (Polymerase Chain Reaction) that would amplify the region with the intended SNP of interest. After one week and after the PCR was run outside of the lab section, the resulting PCR product was purified and treated with restriction enzyme Ahdl in order to prepare for the final analysis of our genotypes. In the third and final week of the project, we analyzed our PCR products by means of agarose gel electrophoresis. By the conclusion of the experiment, we had completed the analysis at the SNP of interest and determined our genotypes for this SNP.
Equation 3.1 can be simplified to the following equation γ(t,m;m_m )= e^(α-βm)/〖(t+c)〗^p (3.2) Where a_0=a+bm_m , α=a_0 ln10 and β=b ln10 are defined. |γ_m (t,m;m_m )|=|∂γ(t,m;m_m )/∂m|=e^α/〖(t+c)〗^p βe^(-βm) (3.3) Where |γ_m (t,m;m_m )| represents the absolute value of the partial derivative of γ(t,m;m_m ), and it is the instantaneous daily rate density of aftershocks of magnitude m at time t following a main-shock of magnitude m_m. e^α/〖(t+c)〗^p denotes the mean instantaneous daily rate of aftershocks at time t following the main-shock of magnitude m_m. βe^(-βm) is the exponential probability density function of aftershock magnitudes.
1.1 Abstract The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample. The purpose of stopped enzyme assay to study B-galactosidase is to determine the effect of temperature and concentrations of substrate on enzyme activity.
The helicase enzyme initiates the DNA process by unwinding the double helix. If it was missing, the DNA would not be able to replicate as the helix structure would not open. The next step of DNA Replication is the binding of RNA primase in the the initiation point of the 3'-5' parent chain. RNA primase can attract RNA nucleotides which bind to the DNA nucleotides of the 3'-5' strand due to the hydrogen bonds between the bases and also provide a starting point for DNA polymerases to extend from. Without it, the DNA nucleotides would not have something to bind to and to start the DNA synthesis.
The hypothesis of the “The effects of a high protein diet on indices of health and body composition – a crossover trial in resistance-trained men” is that the increasing one’s protein intake would have underlying effects on health, performance, and body composition in young males with extensive resistance training experience. In this experiment they are testing the effects of a high protein diet of men that have extensive weight-training experience. The experimental design of this experiment is testing a group of volunteers twice for eight weeks. The first of the eight weeks the subjects consumed their current every diet. The second of the eight weeks of the experiment the subjects consumed a high caloric diet.
No Media mg / ml 1 0.1 N HCl 249.119 2 pH 4.5 Acetate buffer 258.623 3 pH 6.8 Phosphate buffer 260.877 4 Purified water 253.200 *Average of three determinants 8.1.3 UV-VIS spectrophotometric method for OXB 8.1.3.1 Selection of
[14] They exert their antimicrobial effect by inhibiting microbial protein synthesis. In the present study the results showed statistically significant reduction in pocket probing depth (PPD) from baseline to 6 months in both the Test and the Control groups. (p=0.002). Similarly , in both the Test and the Control groups there was a statistical
Biochemical tests are the tests used for the identification of bacterial species based on the differences in the biochemical activities of different bacteria. Bacterial physiology differs from one species to the other. These differences in carbohydrate metabolism, protein metabolism, fat metabolism, production of certain enzymes and ability to utilize a particular compound help them to be identified by the biochemical tests. Gram’s stain was originally devised by histologist Hans Christian Gram in 1884. Gram-positive bacteria stain purple, while Gram-negative bacteria stain pink when subjected to Gram staining.