Electrophoresis – An Introduction • An analytical technique in which the motion of scattered particles run through a fluid under the influence of uniformly charged field is called electrophoresis. • This phenomenon was first observed by Ferdinand Frederic Reuss followed by Arne Tiselius who won the Nobel Prize in chemistry for his research on electrophoresis, adsorption analysis and his discoveries concerning the complex nature of serum protein. It is a technique used in laboratories in order to separate macromolecules based on molecular size and charge. • This technique applies a negative charge so proteins move towards a positive charge. This is used for both nucleic acids and proteins.
After a gram stain was done unknown #257 was identified as a gram positive organism because when observed under the microscope the organism appeared purple with cocci in clusters. The organism was also catalase positive which means that it produced enzyme catalase and bubbled when hydrogen peroxide was added to it. Three test were conducted based on the result of the gram staining procedure. Blood agar with a Novobiocin disk was chosen as well as DNase (DNA) and Mannitol Salts (MSA) agar. The Blood agar is a bright red, opaque plate and the streaking or the inoculation technique was a modified streaking for isolation with a heavy quadrant one.
The latter would require gel electrophoresis. This method uses RFLPs as they travel across a gel, attracted to the other side of the gel with a positive charge. What are restriction enzymes? A restriction enzyme is an enzyme that cuts DNA at only certain (but can be palindromic) sequences in the DNA. These fragments become
The treatment takes 30 minutes to 1 hour and you can resume your normal activities without any problem. Vanquish won’t interrupt your work or daily activity. Four treatments are recommended for best results. Benefits of Vanquish One of the best things about Vanquish is that it has minimal side effects. This treatment procedure also covers larger areas than other methods.
Agarose gel electrophoresis is an easy and common technique of separating and analyzing DNA. The main objective of this lab is to find the sire of the offspring using gel electrophoresis. Gel electrophoresis is used in laboratories to isolate charged molecules like DNA, RNA, and particular proteins according to their specific size. The charged molecules travel through the gel when an electric current is spread across it. The electric current is applied across the gel so that the ends of the gel have a positive charge and the other end has a negative charge.
Introduction In virology, a quantitative assay is a method used to measure the number of virus particles in an inoculated substance. There are various types of quantitative viral assays such as a plaque assay, infectious center assay, pock assay, and transformation assay. All of these assays contain different characteristics and are used to quantify different viral properties. In this lab, the quantitative assay that was constructed was a plaque assay. A plaque assay is a modification of a bacteriophage assay and is used to estimate the titer (concentration of a solution) of a phage stock.
The absorbance of this red pigment; betanin was tested in this lab in relation to the membrane permeability of the beet plant. For a cell membrane to be known as permeable, it means that it has the ability to let a fluid or liquid or even gas to pass through it. A cell membrane could be selectively permeable if it only allows certain molecules to pass through it by a process known as active transport. This process requires energy to move the molecules through the cell. This is why it is important to study how certain concentrations affect the permeability of the cell in question because it leads back to the process of osmosis.
This method is based an Antibody antigen interaction. The Indirect method involves the use of two antibodies to detect the antigenic protein lysozyme. The lysozyme is added to a 96 well plate to coat the plastic and a primary antibody is then added which recognizes the antigen. The primary antibody used was from a different species to that of the antigen it would capture and it referred to as rabbit-anti-lysozyme antibody. The secondary antibody would the overlap the primary antibody by binding to it.
Effect of substrate concentration on enzyme activity Exploration: Introduction: Catalase is an enzyme normally found in many plant and animal tissues. Its purpose is to destroy toxic substances like hydrogen peroxide which is a byproduct in many cellular reactions. In this lab, we will use a catalase solution from yeast and determine the effect of substrate concentration on the action of this enzyme. The substrate of the enzyme will be different concentrations of hydrogen peroxide (H2O2). Catalase works by the following mechanism : 2 H2O2 ------------------> 2 H2O+ O2 Hypothesis: The hypothesis for this experiment is that the foam of O2 produced from the reaction between hydrogen peroxide and catalase will increase in height when the concentration of hydrogen peroxide increases.
It is an analytical method where in a protein sample is electrophoresis on an SDS- PAGE and electro transferred on the nitrocellulose membrane. The transferred protein is detected using specific primary enzymes labeled antibody. Antibodies bind to specific sequences of amino acids, known as the epitope. Because amino acid sequences are different from protein to protein, antibodies can recognize specific proteins among a group of many. Therefore, a single protein can be identified in a cell lysate that contains thousands of different proteins and its abundance quantified through western blot
After receiving an unknown mixture, the sample was streaked for isolation onto TSA, blood agar, and MacConkey plates. Each plate serves as a first step to identify the unknowns. The TSA (tryptic soy agar) can be used to do a gram stain, which differentiates gram-negatives from gram-positives, based on the structural make up of the cell wall (Carson, 2015). The blood agar plate is used to test for hemolytic activity, which is useful for distinguishing gram-positives. A MacConkey plate is selective by inhibiting the growth of gram-positives and differential due to the fermentation of lactose by certain gram-negative species.
The repressor is a regulatory protein that binds to the operator and blocks transcription of the genes of an operon. Inducers bind to the repressors and they also regulate gene expression. In the process of identifying the three strains of E.coli, ONPG (ortho-nitrophenyl b-D galactoside) was used as an indicator. ONPG is a substrate that can detect B-galactosidase, and when it does, it turns yellow. Sarkosyl was also a detergent used in the lab to lyse open cells.
Collecting small fractions is essential in column chromatography because they can be combined together; large fractions can lead to multiple compounds in a specific fraction. The purpose of this experiment was to isolate the three components of Excedrin using column chromatography. Thin layer chromatography (TLC) was used to determine when each of the components had been fully eluted from the column. If there was an overlap in fractions between two components, liquid- liquid extraction was done to separate them. The compounds were characterized via NMR instrumentation and the percent recovery for each compound was calculated to determine whether the isolation was
Narrowing down the unknown microorganism to gram negative, this approach was helpful to take the next step, in some bacteria the cell wall is surrounded by cell enveloped called capsule, also some bacteria make capsule when faced in a harsh environment to protect them. A capsule stain was preform, the results were analyzed and observed. An additional procedure that was done, was the Fast Actin staining which helps to see if the bacteria contains Mycolic acid in their cell walls, which determines the structure and function of the cytoskeleton in living and fixed cells (Shah). As expected for both E.coli and K. Pnenumia the fast acting results were negative. For both E.coli and K. Pnenumia the Oxidase test was positive a reaction was obtained.