Gel Electrophoresis Lab Report

The homogenates provided were made by homogenizing tissues in a sucrose phosphate buffer in a 1:20 ratio. The protein concentration in bovine cells was measured by diluting the homogenate with a 1:5 ratio; 50 microliters of homogenate and 200 microliters of water. Then 5 known protein concentration samples which were 0.4, 0.8, 1.2, 1.6, 2.0 mg/ml of bovine serum were used to determine absorbance with a spectrophotometer. Two additional samples were made; one was blank and the other was for the specific homogenate sample. Then 3 microliters of bradford assay reagent, which indicates the amount of protein present

This experiment’s purpose is to identify and separate DNA macromolecules with a homemade gel and electrophoresis chamber. After separating DNA with a homemade gel and electrophoresis chamber, it must be compared to the regular gel electrophoresis chamber. In order to create a homemade gel electrophoresis chamber, five 9-volt batteries were connected and attached to stainless steel wire which were submerged in the 20x buffer as well as the gel. The two wires acted as the positive and negative electrodes as seen in a gel electrophoresis chamber. After the experiment was done, the homemade gel electrophoresis chamber was proven to mock the regularly used electrophoresis chamber. The experiment suggested that the results would not change if the

In the lab, we were introduced to six different DNA samples, the Crime Scene and 5 suspects. Our Independent variable of the experiment was the restriction enzyme, and our dependent variable was the DNA sample that matched the Crime Scene DNA. With our variables stated. This splices up the DNA into segments that are ready to begin gel electrophoresis.

You do not need to complete this graph if you attach the printout of the computer generated graph

In this experiment, we tested our hypothesis through the use of a PCR and gel electrophoresis. To begin this experiment, we gathered the materials needed. We gathered a screw cap tube that contained the instagene matrix, a 1.5 ml tube, and a small cup containing 0.9% saline solution, P-20 micropipette, P-1000 micropipette, pipet tips, PCR tubes, cap-less microtubes, Master Mix, and water baths. Next, I rinsed my mouth thoroughly for 30 seconds with the saline solution and spit it back into the cup. I then set my P-1000 micropipette to 1,000 μl and transferred 1 ml of my rinse into the test tube. Then, I used the centrifuge to spin the tube at full speed for 2 minutes. I did not have a large enough pellet, so I centrifuged it again for another two minutes. I poured off the supernatant, but I left a small amount of saline at the

I inoculated an Eosin-Methylene Blue (EMB) agar plate and a MacConkey agar plate using Alderson’s streak plate technique (Alderson, 2015, pp. 64-65). Next, using Alderson’s procedure for aseptic techniques for inoculation (Alderson, 2015, p. 53) along with the procedures for each individual test I conducted the biochemical tests on 1 lactose Durham broth (Alderson, 2015, p. 267), 1 glucose Durham broth (Alderson, 2015, p. 267), 1 TSIA slant (Alderson, 2015, p. 267), 1 SIM agar (Alderson, 2015, p. 268), 2 MRVP broths (Alderson, 2015, p. 268) and 1 citrate agar (Alderson, 2015, p. 268). I also tested my microbe’s ability to produce oxidase (Alderson, 2015, p. 267). Finally, I tested for the presence of catalase (Aryal, 2015, para.

Agarose gel electrophoreses is used to separate DNA molecules based on size and molecular weight. The Agarose gel is capable of separating DNA molecules as small as 500 base pairs. This separation is possible due to the negative charged phosphates in the backbone of DNA. The electrical current passing through the gel causes the negatively charged molecules to travel through the gel, which separates the molecules based on size. Smaller and highly negative molecules travel faster through the gel than larger and highly positive molecules.

Agarose gel electrophoresis (AGE) is based upon the simple yet imperative concept that precedes all of biology, as well as every other science – opposites attract. DNA molecules contain phosphate groups that create a negative charge [1]. An electric field across the agarose gel will cause the DNA fragments to migrate to the positive side, allowing visualization of the positions and intensities of various bands [1].

The purpose of this study was to study the plasma membranes of erythrocytes of the human and determine how the majority of polypeptides work with it and where they are located with respect to it. The purification of the erythrocytes was executed by osmotic hemolysis and were washed to the point of being “ghosts”. The polypeptides were then analyzed on the basis of mass by polyacrylamide gel electrophoresis; sodium dodecyl sulfate (SDS) was used to denature the proteins and separate them from the lipids. Furthermore, the location of the membrane proteins were determined by altering the environment around the membrane and recording how they responded. The major conclusion obtained from this study was that the distribution of proteins along the inside and outside of the plasma membranes of erythrocytes exhibits absolute asymmetry.

Implementation of Gel Testing for Antibody Screening and Identification in a Community Hospital, a 3-Year Experience

Supernatant from samples were then mixed in a 1 to 1 ratio with dark blue electrophoresis loading buffer (500mM Tris, 3.84M Glycine, 15% glycerol, 0.25% bromphenol blue). 15ul of each sample was then loaded into agarose gel well. The other wells contained 15ul of Hemoglobin-Albumin, 15ul of Cytochrome C, 5ul of HRP-Basic, and 5ul of HRP-mixture standards.

Gel electrophoresis has had a very big impact on the world of science today. In this present day, scientists now have a simple and definite way to test and contrast a wide variety of different DNA samples, leading to more complex and particular DNA testing (LinkedIn Corporation, 2017). Gel electrophoresis is now a scientific method in which is an absolute necessity in science, particularly in the forensics field. This process of gel electrophoresis gives forensic scientists the ability to retrieve an indisputable link to a person of suspect or anybody for that matter.

Bag #4 had a 40% solution of sucrose and was red. Bag #5 was half filled with pure water with no coloring. Bags 1-5, where all half full with the described solutions, then tied close. Also, another two bags (bag #6 and #7), one containing water (bag #7) and bag #6 with 40% sucrose solution with red food coloring, are completely filled when tied, unlike bags #1-5. The dialysis tubes where knotted at both ends with a simple knot and tested to ensure no leaks, then a 6 inch long string was tied to one end of the bags to aid in retrieval from the beakers. One beaker was filled with enough water to cover all 5 half full bags (bags 1-5), whereas the other was filled with enough 40% sucrose solution to emerge the 2 full bags (bags 6 & 7). Please note that a control bag was present in both beakers, bag #5 in Beaker with water and bag #7 in the Beaker with 40% sucrose solution. At the beginning of the experiment all packets where weighed and recording in grams on the scale, all bags where then placed into their respective beakers. They were weighed at 15 minute intervals, for a total of 60 minutes. Upon weighing all the packets where rinsed and dried thoroughly to prevent any additional solution or water to cling to the exterior of the bags, the results were recorded in Figure 1.

Gel electrophoresis is a routine laboratory procedure in biochemical studies that takes advantage of a protein’s amphoteric nature to determine its molecular weight and charge by running the sample through a gel matrix under the influence of an electrically charged field. A popular example of gel electrophoresis is Sodium dodecyl sulfate-polyacrylamide electrophoresis or SDS- PAGE which has been used in this experiment to supposedly determine albumin and casein’s molecular weights respectively. The system used in gel electrophoresis consists of 3 major parts: a stationary phase, a mobile phase) and an electrical power supply.

The protein-bound dye is then solubilized by weak base for spectrophotometry. This colorimetric assay can be used to estimate the cell number indirectly by providing a sensitive index of total cellular protein content which is nearly related to cell density [12].