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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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 this desired sequence. Once the annealing process is complete‚ the primers are extended (at 30o C) by a DNA polymerase added to the reaction (Mullis et. al
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DNA DIGESTION AND ELECTROPHORESIS In this experiment we will be doing a process called as DNA digestion or also known as restriction digest. A restriction digest is a procedure used in molecular biology to prepare DNA for analysis or other processing. It is sometimes termed DNA fragmentation‚ scientists Hartl and Jones describe it this way: This enzymatic technique can be used for cleaving DNA molecules at specific sites‚ ensuring that all DNA fragments that contain a particular sequence have the
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Enzymes and Electrophoresis Bhumik Patel Phillips 1/16/11 Restriction enzymes are tools in DNA research that can cut DNA into exactly needed pieces. Certain cuts can be rough‚ while others can be clean. Certain cuts can have an organized pattern to have a staggered cut. Other cuts will leave complementary bases with them. Electrophoresis allows the manipulation of DNA to separate and organize those parts. Electrophoresis is the substrate electric movement of the separation of DNA. Gel Electrophoresis
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The team of forensic anthropologists ran a gel electrophoresis with DNA from Skeleton 3 and two missing persons‚ Julia Ly and Teresa Chen to help in DNA identification. This process would allow restriction enzymes to cut by a specific restriction site and run through the gel‚ where the DNA fragments would move from the negative side to the positive side of the gel due to the negative charge of the phosphate group in DNA. The smaller the DNA fragments‚ the further they move down the gel. As mentioned
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Extraction of Bacteria Plasmid DNA and Analysis of extracted DNA Samples Objectives: 1) To study and understand the steps for extract bacteria plasmid DNA. 2) To measure the concentration and purity of extracted DNA by using spectrometric method and agarose gel electrophoresis method. 3) Determine the size of extracted DNA by using agarose gel electrophoresis method. Materials and Methods: (Refer to UDEE2124 lab manual from page 7 to page 10)
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week – we talked about PCR and how important it is in the forensic setting. We will finish off talking about PCR‚ and then we will discuss how it can be used. If we go back to the slide of the double stranded DNA‚ and if we take that to a high temperature‚ the two strands separate‚ you then add the primers‚ which interact with ? On the strand‚ synthesis takes place in the 5-3 direction‚ then you end up with 2 molecules identical to the DNA‚ and then you do another round‚ so it’s an exponential increase
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The Amplification DNA Extraction from minced meat samples using the Polymerase Chain Reaction (PCR) and Gel Electrophoresis for Purification of the DNA. Date: 14th/21st of October 2016 Partner(s): Aisling Loughman. Aim: The aim of the experiment is learn the technique to extract DNA using minced meat samples (Pork‚ Beef and mixed)‚ amplify the extracted DNA using the PCR Technique and further visualise the extracted DNA by Gel Electrophoresis under UV light. Introduction: “The method
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DNA FINGERPRINTING LAB REPORT DNA contains genetic material and information that makes up each individual trait. Every person can be identified by providing his or her genetic information based on a particular DNA strand. DNA information is an effective way of identifying persons if it is used properly. It is used to identify humans in different situations such as crime scenes‚ accident scenes‚ paternity testing‚ soldier remain identification‚ inheritance claims‚ missing person investigations‚
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main source of evidence. Notoriously unreliable‚ these techniques have since faded away to the stunning reliability of DNA forensics. In 1984‚ British geneticist Alec Jeffreys of the University of Leicester discovered an interesting new marker in the human genome. Most DNA information is the same in every human‚ but the junk code between genes is unique to every person. Junk DNA used for investigative purposes can be found in blood‚ saliva‚ perspiration‚ sexual fluid‚ skin tissue‚ bone marrow‚ dental
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