...Agarose Gel Electrophoresis for the Separation of DNA Fragments Pei Yun Lee, John Costumbrado, Chih-Yuan Hsu, Yong Hoon Kim Department of Molecular, Cell, and Developmental Biology, University of California Los Angeles Video Article Chapters 0:05 Title 1:31 Preparation of the Gel 3:21 Setting up Gel Apparatus and Separating DNA Fragments 4:55 Observing Separated DNA Fragments 5:43 Results: Agarose Gel Electrophoresis of PCR Products 6:23 Conclusion Cite this Article Lee, P. Y., Costumbrado, J., Hsu, C. Y., Kim, Y. H. Agarose Gel Electrophoresis for the Separation of DNA Fragments. J. Vis. Exp. (62), e3923, doi:10.3791/3923 (2012). Abstract Agarose gel electrophoresis is the most effective way of separating DNA fragments of varying sizes ranging from 100 bp to 25 kb1. Agarose is isolated from the seaweed genera Gelidium and Gracilaria, and consists of repeated agarobiose (L- and D-galactose) subunits2. During gelation, agarose polymers associate non-covalently and form a network of bundles whose pore sizes determine a gel's molecular sieving properties. The use of agarose gel electrophoresis revolutionized the separation of DNA. Prior to the adoption of agarose gels, DNA was primarily separated using sucrose density gradient centrifugation, which only provided an approximation of size. To separate DNA using agarose gel electrophoresis, the DNA is loaded into pre-cast wells in the gel and a current applied. The phosphate backbone of the DNA (and RNA) molecule...
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...which are also described here. Specific strategies for testing are also discussed. THE BASICS Isolation of DNA and Polymerase Chain Reaction (PCR) DNA or RNA for genetic testing is almost always isolated from peripheral-blood leukocytes. This requires that the blood be drawn in tubes containing some sort of anticoagulant. The preferred anticoagulants are either citrate or EDTA. The cells are lysed followed by removal of the other cellular components and precipitation of the DNA or RNA in ethanol. One of the drawbacks of this approach is that the blood must be rapidly transported to the laboratory where the testing will be performed before the nucleic acids begin to degrade. Recent developments in filter paper technology show promise in obviating this problem. Special filter papers...
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...Experimental Procedures Larval Studies The larvae were acquired from the Honey Bee Research Center located near the University of Guelph. Prior to obtaining the larvae, two 24-well plates were prepared with 300 µL of royal jelly solution. This mixture contained 43.2 mL of Milli-Q water, 14.4 g of lyophilized royal jelly, 1.8 g of glucose, 1.8 g of fructose and 0.6 g of yeast extract. It was stirred in a suitable sized beaker for approximately sixty minutes. Then it was poured into screw-cap veils once it reached a smooth consistency and stored at 4°C until needed. The grafting process consisted of obtaining a frame from one of the hives with larvae at the Honey Bee Research Center. In order to protect the larvae from environmental factors, the frame was wrapped with a towel before the grafting process ensued. With a small grafting tool, the larvae were individually scooped out of the comb on the frame. The larvae were placed into one of the each wells on the twenty-four well plate to float on the royal jelly solution (Figure 9). This was conducted cautiously to ensure that the larva did not flip over since oxygen intake occurs only on one its sides. Once the twenty-four well place...
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...Gene Marker Identification Targeting Toll-Like Receptor 4 (TLR4), Breast Cancer 1 (BRCA1), and Adenosine Triphosphatase 1 Alpha 1 (ATP1A1) Genes: Assessing Their Association With Subclinical Mastitis Cases in Dairy Water Buffaloes, Bubalus bubalis Thesis Proposal Cyndi Candelaria Biendima Patricia Malapit Cabatit Submitted to the Department of Biology College of Arts and Sciences University of the Philippines Manila Padre Faura, Ermita, Manila In partial fulfilment of the requirements for Undergraduate Thesis (BIO 200) TABLE OF CONTENTS Title Page1 Table of Contents2 Introduction3 Review of Related Literature6 Proposed Methodology14 Presentation of Results20 Literature Cited22 Line Item Budget26 Project Timeline27 1.0 INTRODUCTION 1.1 Background of the Study Cases of intramammary infections such as mastitis in water buffaloes contribute to large annual losses in milk production and net profit for smallholder farmers in the Philippines. Social and economic factors might prevent households from diagnosing, treating, and eliminating from circulation those animals or animal products, such as milk, that are afflicted with mastitis or which came from individuals afflicted with mastitis; this is especially true in the case of the asymptomatic subclinical mastitis, which tends to become chronic and difficult to eradicate by conventional antimicrobial therapies (Brouillette & Malouin, 2005; Ng et al., 2010). With the advent of technology...
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...evolution by Joseph Yang April 13, 2015 by Joseph Yang April 13, 2015 Evolution is a change in the gene frequency in a population through time. Evolution causes an organism’s ability to survive to increase. In many cases, predators evolve to catch their prey more efficiently. Evolution is not one sided. The prey also evolves to avoid predation. In this paper, I will discuss three researches done on a prey’s evolution to survive against predation. I will discuss the tail autonomy in Tokay gecko (Sanggaard et al. 2012), the odorous and non-fatal secretion in Wrinkled frog (Yoshimura and Kasuya 2013), and the avoidance response in Brownbanded bamboo shark embryos (Kempster et al. 2013). Together, these studies show the effectiveness and vital impact of evolution to the survival of prey. Sanggaard et al. (2012) researched evolution in Tokay gecko (Gekko gecko) to investigate the mechanism of their tail autonomy by facilitating autonomy. The geckos were euthanized by pentobarbital and the tails were removed by induced autonomy. Moisture from the exposed end of the tail was collected and subjected to Gel Electrophoresis and Mass Spectrometry and analyzed using MS-BLAST. The tails’ structure was analyzed through multiple methods. Through Magnetic Resonance Imaging, a Micro Imaging 5 probe, a Great 60 Imaging system, a saddle coil, and a gradient cooling temperature of 20°C was utilized. Electron Microscopy and 3D Fast Low-Angle Shot sequence were utilized. Scanning Electron Microscopy...
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...* DNA technology – Methods for studying and manipulating genetic material, has rapidly revolutionized the field of forensics, the scientific analysis of evidence for legal investigators * Has practical applications beyond its use in forensic science * Include the use of gene cloning in the production of medical and industrial products, the development of genetically modified organisms for agriculture, and even the investigation of genealogical questions * DNA evidence was used to solve a double murder in England * Showed that two murders could have been committed by the same person * Showed the innocence of someone who confessed to one of the murders * Showed the absence of a match in 5,000 men tested when the murderer persuaded another man to donate blood in his name * Showed a match with the murder and DNA found with both victims * Biotechnology: The manipulation of organisms or their components to make useful products * Grew out of discoveries made about 60 years ago by American geneticists Joshua Lederberg and Edward Tatum * Performed a series of experiments with E.coli that demonstrated that two individual bacteria can combine genes * Genetic engineering involves manipulating genes for practical purposes * Gene cloning leads to the production of multiple identical copies of a gene-carrying piece of DNA * Recombinant DNA is formed by joining DNA sequences from two different sources ...
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...INTRODUCTION: Bangladesh (Mangifera indica L.) belongs to the family Anacardiaceae, is an important and popular fruit of Bangladesh. It has a unique position in respect of nutritional quality, taste, consumer’s preference etc., among the fifty kinds of fruits grown in Bangladesh (Ahmad, 1985). The fruit is believed to have originated in the Eastern India, Asam, Burma or in the Malayan region (Mukherjee, 1997). Mango is now recognized as one of the choicest fruits in the world market for its excellent flavour, attractive color and delicious taste. It has medium calorific and high nutritional values. Carbohydrate content in ripe mango pulp is 16.9% (Salunkhe and Desai, 1984). Besides, mango contains appreciable quantity of provitamin A, vitamin C and soluble sugar (Samad et al., 1975). The unripe fruits contain nearly 50% more vitamin C than the ripe ones and in mineral content, mango holds an average position among fruits and in containing iron, unripe mango is the first and ripe fruit, about the 16th position among all major fruits (Hossain, 1989). The fruit has really of immense value in respect of money and prosperity. In Bangladesh it is called as “King of the fruit”. Due to certain limitations of soil and climate conditions, the mango grows better in some selective areas of Bangladesh. The leading mango growing districts are Nawabgonj, Rajshahi, Rangpur, Dinajpur and Kushtia. It is grown over wide geographical areas particularly in India, Pakistan, Brazil, Mexico, the Philippines...
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... Guidance for this Assignment : | Date set: Task 1 27/1/14 | Date set: Task 2 10/2/14 | Date set: Task 3 24/2/14 | Date set: Task 4 10/3/14 | Remember! | * Your attention is drawn to the comments made in the Course Handbook with regard to both the completion and submission of work * Assessment deadlines must be met * Any student found guilty of cheating or plagiarism may be withdrawn from the unit at the discretion of the Assessment Board. | Internal verification: | Date: | | Name | | | | Signature | | Aim and purpose:-To develop understanding of the principles of Mendelian genetics and to develop knowledge and practical techniques used in commercial, analytical and research laboratories | | GRADING CRITERIA To achieve a pass grade the evidence must show that the learner is able to: | To achieve a Merit grade the evidence must show that the learner is able to: | To achieve a distinction grade the evidence must show that the learner is able to: | P1 Compare and contrast the structure of various nucleic acids. | M1 Explain how genetic information can be stored in a sequence of nitrogenous bases in DNA. | D1 Explain the steps involved in biosynthesis of protein including the roles of...
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...nutrition, and the environment. I studied some reports and papers that focus on the environmental impact analysis, such as Life Cycle Analysis (LCA). I also came across some papers by Prof. Mathys and colleagues on the LCA of meat alternatives and study on the insect as an emerging sustainable food source. Furthermore, the focus of the Sustainable Food Processing group is fit right into my study interests. A collaboration with Prof. Mathys and the team would be a great opportunity for me to increase my research skills and also to learn more about the food sustainability area from the experts. My education and professional experience have aided me to prepare myself for a graduate study. Last year, I had an opportunity to do a summer research to develop a method to quantify glutamine in barley grains. The project was very challenging because glutamine is not an easy substance to be quantified due to its sensitivity. The research was divided into some stages of experiments. The first stage was to do trials of existing protein extraction methods, followed by protein content analysis by using protein analyser and then gel electrophoresis analysis to confirm whether prolamins (i.e. protein fraction that is rich in glutamine) was extracted. The second stage was the derivatization step to transform glutamine into a stable acid, and the last stage was hydrolysis and amino acid profile analysis by using HPLC. The first stage of the research was succeeded, there were two methods that resulted...
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...Molecular analysis of Jawahar Rice Hybrids for genetic purity and fingerprinting Amrita Ingole, Abstract Varietal identification and genetic purity has attained critical importance worldwide especially in the context of plant variety protection. The estimation of hybrid seed purity is done conventionally by grow out test, which is based on the assessment of morphological and floral traits in plants grown to maturity, which locks up the capital while awaiting the results for GOT. GOT could be replaced with DNA based assays. The present investigation was undertaken with an objective to identify distinguishable microsatellite markers to establish fingerprinting of rice (Oryza sativa L.) hybrids, assessing variation within parental lines and testing the genetic purity of hybrid seed develpod by JNKVV Jabalpur, 4 cms, 4 restores and 5 hybrids were employed in this study. About 52 most informative microsatellite markers were employed for fingerprinting five rice hybrids and their parental lines. A total of 77 alleles were detected, and the number of alleles per locus ranged from 1 to 6, with an average of 2.56±1.33 primer pair. Eleven markers amplified specific alleles those separated the male from female lines. 13 markers were found polymorphic for female lines. Marker RM 164 clearly differentiates all the 4 CMS lines under study. 24 markers were found polymorphic among the male parents. No such marker was found that could differentiate all the male lines from each other...
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...Introduction IgG is an antibody probe that has an affinity for binding specifically to the IgG protein antigen. The research goal was to identify how closely the IgG of the tested species are related. This was accomplished by examining how goat IgG, specific for goat protein, responded to the same protein from cows, pigs, rats, and humans, and it was determined which animals have the most similar antigen binding reactions. The variability in each animal’s IgG protein was revealed by the level of affinity that the anti-goat IgG probe had for the species tested. Since the variability in each animal’s proteins is due to their genetic coding sequence this study clarified which species have the most similar genetic code. Immunoglobulins are the antigen-recognition molecules of B cells and are used as the main effector function in adaptive immunity. (Janeways, 2001) Their are five major immunoglobulin classes: IgA, IgD, IgE, IgG, and IgM and all five Ig classes are present in mammals and are produced from B lymphocytes as part of the immune response system. (Urich, 1994) IgG antibodies are large molecules, having a total molecular weight of 150kDa, composed of two heavy (H) peptide chains weighing approximately 50kDa and two light (L) peptide chains approximately 25kDa. (Janeways, 2001). The region of Light and Heavy chains connected by a disulfide bond makes up the Fragment antigen binding (Fab) structure, while the remaining Heavy chain region is referred to as the...
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...experiments in this paper done through instruction in graduate course: Biotechnology laboratory in the natural sciences and mathematics department at the university of texas at dallas, RICHARDSON, TX 75080 march 2016 [Company name] | [Company address] ------------------------------------------------- γ-globin analysis by expression profiling through RT-qPCR, quantification through ELISA, and oxidative stress management analysis by FACS from KU812F cells under treatment by δ aminolevulinic acid, succinylacetone, and N-methyl mesoporphyrin Shaan Sarode, Jose Cordero, and Dr. Li Liu experiments in this paper done through instruction in graduate course: Biotechnology laboratory in the natural sciences and mathematics department at the university of texas at dallas, RICHARDSON, TX 75080 march 2016 [Company name] | [Company address] ------------------------------------------------- γ-globin analysis by expression profiling through RT-qPCR, quantification through ELISA, and oxidative stress management analysis by FACS from KU812F cells under treatment by δ aminolevulinic acid, succinylacetone, and N-methyl mesoporphyrin Shaan Sarode, Jose Cordero, and Dr. Li Liu ABSTRACT Hemoglobinopathies refer to a group of blood related disorders that encompass important disease such as thalassemia and sickle cell disease. Because many of these disease are hereditary more aggressive genetic therapies are showing promise as possible avenues of treatment. One such method is to re-express...
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...Biological Sciences Impact Factor ISSN 1449-2288 News feeds of IJBS published articles Manuscript Status/Login Contact 6 May 2015 Home Index & Ranking Current Issue Archive Cover Images Editorial Board Author Info Submission Special Issues Contact Journal of Genomics now in PubMed/PubMed Central. Submit manuscript... Top Introduction Materials and Methods Results Discussion Supplementary Material Acknowledgements References International Journal of Medical Sciences Journal of Cancer Theranostics Journal of Genomics PubMed Central Indexed in Journal Impact Factor PDF Int J Biol Sci 2015; 11(5):546-558. doi:10.7150/ijbs.11084 Research Paper Evidence for Fungal Infection in Cerebrospinal Fluid and Brain Tissue from Patients with Amyotrophic Lateral Sclerosis Ruth Alonso1, Diana Pisa1, Ana Isabel Marina1, Esperanza Morato1, Alberto Rábano2, Izaskun Rodal2, Luis Carrasco1 Corresponding address 1. Centro de Biología Molecular “Severo Ochoa”. c/Nicolás Cabrera, 1. Universidad Autónoma de Madrid. Cantoblanco. 28049 Madrid. Spain. 2. Department of Neuropathology and Tissue Bank, Unidad de Investigación Proyecto Alzheimer, Fundación CIEN, Instituto de Salud Carlos III, Madrid. Spain. How to cite this article: Alonso R, Pisa D, Marina AI, Morato E, Rábano A, Rodal I, Carrasco L. Evidence for Fungal Infection in Cerebrospinal Fluid and Brain Tissue from Patients...
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...ANALYSIS OF FOOD PRODUCTS FOOD SCIENCE 581 Instructor Prof. D. Julian McClements Room 238, Chenoweth Lab Tel: 545 1019 Email: mcclements@foodsci.umass.edu Course Description Food products are analyzed for a variety of reasons, e.g., compliance with legal and labeling requirements, assessment of product quality, determination of nutritive value, detection of adulteration, research and development. The lectures will cover the basic principles of analytical procedures and techniques commonly used to provide information about the chemical composition, structure and physical properties of food materials. The aim of the laboratory classes is to give students experience in performing food analysis experiments, analyzing data and reporting their findings. In addition, students are expected to work in teams on a special project where they will identify and critically assess the most appropriate analytical methods for analyzing the properties of a particular food product. [4 Credits]. Textbooks * Introduction to Food Analysis. S.S. Nielsen, 1998. Aspen Publishers - The best general overview of food analysis techniques currently available. (Required). * Food Analysis: Theory and Practice. Y. Pomeranz and C.E. Meloan, Chapman and Hall - General overview of food analysis techniques (Useful) * Food Analysis: Principles and Techniques. D.W. Gruenwedel and J.R. Whitaker, Marcel Dekker - General overview of food analysis techniques (Useful) * Analytical Chemistry of Foods...
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...Review Paper Inulin - a versatile polysaccharide with multiple pharmaceutical and food chemical uses. Thomas Barclaya, Milena Ginic-Markovica, Peter Cooperb,c, Nikolai Petrovskyc,d a Flinders University, Adelaide, Australia 5042 Cancer Research Laboratory, ANU Medical School at the Canberra Hospital, Australian National University, Canberra, Australia 2605 c Vaxine Pty Ltd, Flinders Medical Centre, Adelaide Australia 5042 d Department of Endocrinology, Flinders Medical Centre, Adelaide, Australia 5042 b Received: 27 August 2010 Accepted: 10 October 2010 ABSTRACT á-D-glucopyranosyl-[â-D-fructofuranosyl](n-1)-D-fructofuranoside, commonly referred to as inulin, is a natural plant-derived polysaccharide with a diverse range of food and pharmaceutical applications. It is used by the food industry as a soluble dietary fibre and fat or sugar replacement, and in the pharmaceutical industry as a stabiliser and excipient. It can also be used as a precursor in the synthesis of a wide range of compounds. New uses for inulin are constantly being discovered, with recent research into its use for slow-release drug delivery. Inulin, when in a particulate form, possesses anti-cancer and immune enhancing properties. Given its increasing importance to industry, this review explains how inulin's unique physico-chemical properties bestow it with many useful pharmaceutical applications. KEY WORDS: Inulin, polysaccharide, fructose, excipient, vaccine, adjuvant INTRODUCTION ...
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