3.1 Source of the Recombinant DNA Polymerase SK72 The recombinant E. coli BL21 (DE3) containing the pET 32b/ DNA polymerase SK72 gene was provided by the Laboratory of Enzyme and Microbial Technology‚ Institute of Bioscience‚ Universiti Putra Malaysia. 3.2 Preparation of the Lysogeny Broth (LB) Agar Plates LB agar plates were prepared by dissolving 10g tryptone‚ 5g yeast extract‚ 5g NaCl and 15g bacterial agar in 950mL of deionized water. The pH of the medium was adjusted to 7.5 using 1M NaOH before
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Objectives Protein Isolation: Protein isolation for a western blot uses detergents and mechanical force to separate seeded cells from their container. Eukaryotic cells are attached to the surface of a flask by cadherins. In the past‚ we’ve separated the cells from the flask by breaking these bonds with a protease‚ but in order to keep the proteins intact‚ a different method needs to be used to extract the proteins. In protein extraction for a western blot‚ we use detergents to lyse the membrane
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agarose gel electrophoresis in an emulation of DNA fingerprinting. The task‚ which was successfully carried out was to determine whether DNA from suspects A‚ B or C matches the sample of blood found at the murder scene (X). The process of PCR acts in the same way as DNA replication but is restricted to specific DNA samples of interest. By amplifying the necessary DNA sequence‚ this procedure is able to produce a usable DNA sample for agarose gel electrophoresis. Agarose gel electrophoresis is a method
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The Isolation‚ purification and identification of Proteins assayed From Bovine Liver Using SDS Gel Electrophoresis‚ Mass Spectroscopy and Western Blotting Abstract The purpose of the experiment was to isolate and recognize varying protein solubilization and assaying methods by using bovine liver protein. The experiment implicated the impact of different types of solvents like ethanol‚ water‚ PBS‚ PBS+1% Triton x-100‚ and PBS+2% SDS on protein solubilization. Bradford and Ghosh/Dumbroff
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and is used to distinguish from different species based on variation‚ commonality‚ or evolutionary divergence. First‚ proteins are extracted from the tissue and loaded into a gel matrix. The matrix will separate the proteins according to size using an electric current. Proteins that are separated after are blotted from the gel and onto a paper membrane. An antibody is then added to the membrane paper and causes a colored reaction. Following the reaction‚ the results help detect and quantify a single
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Unit 7 Topic 2 – Reading 7 Mystery of the Stolen Artifacts Federal and state laws protect archaeological remains on public lands. These laws are important for preserving our national and state heritage. Unfortunately‚ there are people who discover these sites‚ excavate the artifacts‚ and sell them for personal gain. These people are called “pot hunters”. This script is a fictional trial of a Mr. Pete Anderson who was accused of illegally taking archeological artifacts from public land. During a
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METHODS The method used in this lab to map the proteins was the method of Polyacrylamide gel electrophoresis. This method can be used to separate the proteins present in the fish muscle and separates them on size. Due to the fact that they are separated by size‚ the proteins can be compared because similar proteins with stop at the same spot in the gel. So measuring the bands that show up on the gel you can determine if different fish species have similar proteins. The first thing that is
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Ostry‚ Laichmannova‚ Ruprich‚ 2010). Enrico Dainese and his partners did another similar study‚ on soybeans specifically. Like our experiment conducted on the cornbread mix‚ Dainese and his colleagues followed their PCR results with an Agarose Gel Electrophoresis (Dainese‚ Angelucci‚ De Santis‚ Maccarrone and Cozzani‚ 2004). An additional experiment closely related to the one performed by my partners and I is a study done in Brazil to better detect for GMO within their foods sold in markets a other places
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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 based on molecular weight under the influence of an applied electrical field and then used to prepare for the Western Blot (#1 Lehninger). The support medium used is a polyacrylamide gel and‚ it also used sodium
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1. The central purpose of this paper is to describe all of the methods of how PCR was developed and the results of the experiments involving extraction and amplification of DNA (Mullis et. al. 1986). 2. PCR has the ability to isolate specific DNA sequences with the use of primers. This is done by denaturing the DNA (at 95o C) so it is able to anneal to the primers that specify a fragment to be amplified (Mullis et. at. 1986). These primes anneal to a specific sequence of DNA in order to amplify
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