2 ZOOLOGICAL SCIENCE 13: (1996) 1996 Zoological Society of Japan Phylogenetic Position of the Japanese River Otter Lutra nippon Inferred from the Nucleotide Sequence of 224 bp of the Mitochondrial Cytochrome b Gene Tomohiko Suzuki*, Hajime Yuasa and Yoshihiko Machida Department of Biology, Faculty of Science, Kochi University, Kochi 780, Japan ABSTRACT A 224 bp fragment of the mitochondrial cytochrome b gene has been amplified from a 30- year-old mummy-like specimen of the Japanese river otter Lutra nippon by polymerase chain reaction (PCR). The amplified products were subcloned in the Smal site of puc18 and sequenced. The sequence was different from those of the congeneric Eurasian otters Lutra lutra (Latvia) and Lutra lutra (China) in 7-9 nucleotides, all of which were located at the third position of a codon and identified as transitional differences A G or C T. The phylogenetic analysis using the 224 bp sequences of Lutra nippon, Lutra lutra (Latvia), Lutra lutra (China), Aonyx cinerea (Asian small-clawed otter), Mustela sibirica and Mustela itatsi (weasels) supports the recent morphological study that the Japanese river otter is not a subspecies of Lutra lutra, but a distinct species, Lutra nippon. We found that Lutra nippon and Lutra lutra contain the cytochrome b-like sequences, that appear to be a pseudo-form of cytochrome b gene. The sequences are characterized by the presence of deletion and termination codons, by the presence of several types of sequences with minor variations, and by the faster evolutionary rate compared with that of the mitochondrial cytochrome b gene, The genes would present in the nuclear DNA rather than in the mitochondrial DNA, as in the case of the nonfunctional cytochrome b-like sequences previously reported in a rodent. INTRODUCTION The Japanese river otter, a nearly extinct species, has long been thought to be a subspecies whiteleyiofthe Eurasian otter Lutra lutra. Recently, Imaizumi and Yoshiyuki (1989) reexamined the morphological characters in detail, and gave a new species name Lutra nippon, to the Japanese otter from Shikoku and Honshu in Japan. They described that Lutra nippon is a primitive species with relatively longer tail and longer facial portion of cranium than other Eurasian species of Lutra. However, Wilson and Reeder (1993) considered it to be a junior synonym of Lutra lutra. To investigate the possibility for reintroduction of the Eurasian otters to Japan, it is inevitably needed to make clear the phylogenetic position of the Japanese otter. Thus we decided to analyze the sequence of DNA fragment amplified from mummy-like and stuffed specimens of Lutra nippon, under the condition that no living Japanese otters are available. We compare the sequence of Lutra nippon with those of several related taxa, Lutra lutra from Latvia and L. lutra from China (Eurasian otter), Aonyx cinerea (Asian small-clawed otter) and Mustela sibirica (Siberian weasel) and Mustela itatsi (Japanese weasel), and discuss the phylogenetic position of the Japanese river otter. * To whom all correspondence should be addressed. MATERIALS AND METHODS DNA analysis of Lutra nippon DNA was isolated from the muscle (ca. 0.1 g) of a 30-year-old mummy-like specimen deposited at the Ehime Prefectural Museum, Japan, and from the muscle (ca. 0.1 g) of at least 30-year-old stuffed specimen at the Kochi Prefectural Office, Japan of Lutra nippon with a conventional phenol-chloroform method. The DNA was further purified by a spin column of S-400HR (Pharmacia). A 224 bp fragment of mitochondrial cytochrome b gene was amplified for 30 cycles each consisting of 0.5 min at 94 C for denaturation, 0.5 min at 55 C for annealing and 1 min at 72 C for primer extension, using ca. 10 ng of template DNA, by PCR (Saiki et al., 1988). The primers used are: F1: CATCCAACATCTCAGCATGATGAAA (25mer) R4: ATGTTTCATGTTTCGGTGAATATAT (25mer) F1 is the "conserved" primer designed for amplification of mitochondrial cytochrome b gene (Kocher et al., 1989), and R4 was based on the sequence of Mustela itatsi. As an enzyme, Taq DNA polymerase (Promega or AGS GmbH) was used. The amplified products were subcloned in the Smal site of puc18, and sequenced by the dideoxy chain termination method with BcaBEST DNA sequencing kit (Takara) and non-ri Uniplex DNA detection kit (Millipore) (Suzuki and Takagi, 1992). Several clones were also sequenced with a PRISM dye terminator cycle sequencing kit using Model DNA sequencer (Applied BioSystems). DNA analyses of Lutra lutra, Aonyx cinerea, Mustela sibirica and Mustela itatsi DNAs were isolated either from the fresh muscle (Lutra lutra (Oji Zoo, Japan, originated from Latvia), Aonyx cinerea (Noichi Zoo, Japan), Mustela sibirica (obtained from Kochi, Japan) and Mustela itatsi (two individuals, obtained from Fukushima, Japan)) or from fresh
3 622 T. Suzuki, H. Yuasa and Y. Machida hairs (L lutra (Asa Zoo, Japan, originated from China) and L. lutra (Noichi Zoo, imported from Europe)) with a conventional phenolchloroform method. Mustela sibirica and M. itatsi were identified based on their sequences of cytochrome b gene reported by Masuda and Yoshida (1994). The subspecies of Lutra lutra were not considered. The DNA of Lutra lutra (killed by traffic accident in Asahikawa in 1989, Japan, and identified as Lutra lutra, but not Lutra nippon (Tsutsumi, 1990) was kindly supplied by Dr. Toshihiko Shiroishi of the National Institute of Genetics. A 307 bp fragment of mitochondrial cytochrome b gene was amplified by PCR with the "conserved" primers (Kocher
4 Cytochrome b Sequence of Lutra nippon 623 Fig. 1. Comparison of the nucleotide sequences of the mitochondrial cytochrome b and cytochrome b-like sequences of Lutra nippon (Ehime and Kochi) with those of L lutra (Latvia, China, Europe and Asahikawa), Aonyx cinerea, Mustela sibirica and M. itatsi. The sequence length, 224 bp forl..nipponand 307 bp for the others. Several representatives of the cytochrome b-like sequences (ps) are shown. Positions of the deletion and termination codon in the cytochrome b-like sequences are boxed. Euro., Europe; Asahi., Asahikawa. et al.,1989), F1: CATCCAACATCTCAGCATGATGAAA (25mer) R1: CCCCTCAGAATGATATTTGTCCTCA (25mer) The amplification conditions were as for in Lutra nippon. The amplified products were subcloned in the Smal site of puc18, and at least three clones were sequenced. RESULTS AND DISCUSSION We have amplified the 307 bp fragment of the mitochondrial cytochrome b genes from the Eurasian otters Lutra lutra (Latvia), L. lutra (China), L lutra (Europe) and L. lutra (Asahikawa), the Asian small-clawed otter Aonyx cinerea and the weasels Mustela sibirica and Mustela itatsi, subcloned the amplified products in puc18 and determined their sequences. The sequences of L. lutra (Europe) and L. lutra (Asahikawa) agreed completely with those of L lutra (China) and L lutra (Latvia), respectively. The sequences of two individuals of Mustela itatsi were identical, but different in three positions from that reported recently (Masuda and Yoshida, 1994), in the sequence positions (see Fig. 1).
5 624 T. Suzuki, H. Yuasa and Y. Machida On the other hand, many attempts were made to amplify the DNA fragment of cytochrome b gene of Lutra nippon using various year-old samples, such as the liver, heart and skin preserved in formalin, the tanned leather and the hairs of stuffed specimen, but all attempts were unsuccessful. We have succeeded in amplifying the 224 bp fragment of the mitochondrial cytochrome b gene from a 30-year-old mummy-like specimen of Lutra nippon from Ehime (Fig. 1). Direct sequencing of the amplified products suggested the presence of more than two types of sequence. Therefore the products were subcloned in puc18 and 30 clones were sequenced. Among them, nine clones were ambiguously identified to be the mitochondrial cytochrome b sequence of Lutra nippon since the sequence was apparently similar to the mitochondrial sequences from other Lutra species, of which eight clones had the same nucleotide sequence (represented by the sequence of L. nippon (Ehime)-c4 in Fig. 1) but the remaining one (the sequence of L. nippon (Ehime)-c5 in Fig. 1) was different in two positions (21: C T and 92: T A). The eleven clones had the cytochrome b-like sequences as described below. The remaining 10 clones, unexpectedly, had the almost identical sequence to the pig cytochrome b sequence, suggesting some serious DNA contamination occurred during storage of the mummy-like specimen. We found that the DNA from a mummy-like Lutra nippon (Ehime) contains a cytochrome b-like sequence. Surprisingly, all of the amplified products from another Lutra nippon from Kochi (at least 30-year-old stuffed specimen) corresponded to the cytochrome b-like sequences, and no mitochondrial cytochrome b fragment was amplified. In addition, the cytochrome b-likefragments of 307 bp were amplified, with a yield of 30-40% of the total clones, from the DNAs prepared from the fresh liver or hairs of the Eurasian otters L. lutra (Europe and Asahikawa), but not from the DNAs from L lutra (Latvia and China). Thus the presence of cytochrome b-\\ke sequence is not an artifact during a long storage of DNA. The cytochrome b-like sequence are also found in the nuclear DNA in a rodent (Smith et al., 1992). The cytochrome b-like sequences of Lutra are characterized by the presence of a deletion to cause a reading frame shift and termination codons (Fig. 1), by the presence of several types of sequences with minor variations (Fig. 1), and by the apparently faster evolutionary rate compared with the mitochondrial cytochrome b sequence (see Fig. 2). Thus the cytochrome b-like sequence Fig. 2. A phylogenetic tree constructed from the sequence alignment (based on sequence in Fig. 1) of the mitochondrial cytochrome b and cytochrome b-like sequences with the Neighbor-Joining method. The numbers at each branching point show bootstrap values (100 replications), ps represents the cytochrome b-like sequence.
6 Cytochrome b Sequence of Lutra nippon 625 appears not to function and is present as a pseudo-form. We have not examined whether the cytochrome b-like sequence of Lutra is present in the mitochondrial DNA or in the nuclear DNA. The nucleotide sequences of the mitochondrial cytochrome b and cytochrome b-like genes of Lutra nippon are aligned with those of L. lutra (Latvia, China, Europe and Asahikawa), Aonyx cinerea, Mustela sibirica and Mustela itatsi, in Fig. 1. Of the sequence 1 to 224, Lutra nippon (C4) was different from those of L lutra (Latvia) and L. lutra (China) in 9 and 7 nucleotides, respectively, all of which were located at the third position of a codon and identified as transitional differences A G or C T. The difference within Lutra lutra was 4 nucleotides. These nucleotide substitutions cause no amino acid replacement. On the other hand, there are 6 nucleotides difference in the two congeneric weasels Mustela sibirica and Mustela itatsi in this region. Thus the sequence difference between Lutra nippon and Lutra lutra (7-9 nucleotides: 3.6% in average) is larger than that (6 nucleotides) between the two Mustela species. This is also consistent with the idea that in the Mustelidae species, more than 3.5% difference in sequence of cytochrome b gene may correspond to the difference of distinct species (Masuda and Yoshida, 1994). The percentage difference between the nucleotide sequences are shown in Table 1. Figure 2 shows a phylogenetic tree constructed from the sequence data obtained in this study with the Neighbor-Joining method in the PHYLIP package version 3.5c (Felsenstein, 1993). The same topology was reconstructed with the Fitch and Margoliash method (Felsenstein, 1993). The sequences were divided into two main clusters, one is for the mitochondrial cytochrome b genes and the other for cytochrome b-like sequences. As stated above, the cytochrome b-like sequences were amplified from the DNAs of Lutra nippon (2 individuals, from Kochi and Ehime), L. lutra (Europe) and L. lutra (Asahikawa). The branching pattern of the cytochrome b-like sequences of the two Lutra species was the same as in the mitochondrial cytochrome b genes, but the evolutionary rate for the cytochrome b-like sequences was at least two-times faster, in agreement with the general characteristics of pseudogenes. The tree also clearly shows that there is a large genetic difference between Lutra nippon and Lutra lutra, and this result is consistent with the recent morphological study that the Japanese river otter is not a subspecies of the Eurasian otter Lutra lutra, but a distinct species, Lutra nippon (Imaizumi and Yoshiyuki, 1989). However, to get a conclusive evidence for the phylogenetic relationship between the two species, longer DNA sequence of Lutra nippon must be determined. This would be done when the fresh sample of the Japanese river otter was obtained. ACKNOWLEDGMENTS We thank Dr. Serge N. Vinogradov of Wayne State University for reading the manuscript. We also thank Drs. Hitoshi Suzuki, Hidetoshi Tamate, Takahiro Furukohri and Tomikazu Arita for valuable suggestions, Dr. Toshihiko Shiroishi for supplying us the DNA of Lutra lutra (Asahikawa), Shigekazu Suzuki, Wahei Sakuma, Yoshiyuki Suzuki, Naotaka Ohno and the staffs of the Tobe Zoo, Noichi Zoo, Oji zoo, Asa Zoo, Asahiyama Zoo, Ehime Prefectural Museum and Kochi Prefectural Office for supplying specimens used in this study. This work was supported partially by grants from the Environmental Agency, Japan and Kochi University. REFERENCES Felsenstein J (1993) PHYLIP (Phylogeny Inference Package) version 3.5c. Distributed by the author. Department of Genetics, University of Washington, Seattle, U.S.A. Imaizumi Y, Yoshiyuki M (1989) Taxonomic status of the Japanese Table 1. Percentage difference between the nucleotide sequences (224 bp) shown in Fig. 1 A B C D E F G H l J K L M N O P Lat. Asahi. China Euro. Ehime c4 Ehime c5 Ac. M.S. M.i. L.l. Asahi. ps3 Asahi. ps1 Euro. ps34 Euro. ps1 Ehime ps7 Kochi ps7 Kochi ps17 B C D E F G H I J K L M N 0 P Lat., Latvia; Asahi., Asahikawa; Euro., Europe.
7 626 T. Suzuki, H. Yuasa and Y. Machida otter (Carnivora, Mustelidae), with a description of a new species. Bull Natn Sci Mus, Tokyo Ser A 15: Kocher TD, Thomas WK, Meyer A, Edwards SV, Pääbo S, Villablanca FC, Wilson AC (1989) Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proc Natl Acad Sci USA 86: Masuda R, Yoshida MC (1994) A molecular phylogeny of the family Mustelidae (Mammalia, Carnivora), based on comparison of mitochondrial cytochrome b nucleotide sequences. Zool Sci 11: Saiki RK, Gelfand D, Stoffel S, Scharf S, Higuchi R, Horn G, Mullis K, Erlich H (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: Smith MF, Thomas WK, Patton JL (1992) Mitochondrial DNA-like sequence in the nuclear genome of an akodontine rodent. Mol Biol Evol 9: Suzuki T, Takagi T (1992) A myoglobin evolved from indoleamine 2,3-dioxygenase. J Mol Biol 228: Tsutsumi N (1990) A report on the river otter in Asahikawa (in Japanese). Department of Health and Environment, Hokkaido Government Wilson DE, Reeder DM (1993) Mammal Species of the World - A Taxonomic and Geographic Reference 2nd ed. Smithon Inst Press, Washington and New York (Received February 28, 1996 / Accepted April 17, 1996)
THE POLYMERASE CHAIN REACTION (PCR) OBTECTIVE: To amplify a region of ribosomal DNA from a small amount of genomic DNA. INTRODUCTION: The polymerase chain reaction (PCR) was developed in 1983 by Kary Mullis,
Using the NCBI Genome Databases to Compare the Genes for Human & Chimpanzee Beta Hemoglobin Author(s) :Susan Offner Source: The American Biology Teacher, 72(4):252-256. 2010. Published By: National Association
Maximum-Likelihood Estimation of Phylogeny from DNA Sequences When Substitution Rates Differ over Sites1 Ziheng Yang Department of Animal Science, Beijing Agricultural University Felsenstein s maximum-likelihood
Biological Sciences Initiative HHMI PCR Polymerase Chain Reaction PCR is an extremely powerful technique used to amplify any specific piece of DNA of interest. The DNA of interest is selectively amplified
Cat. # R010A For Research Use PrimeSTAR HS DNA Polymerase Product Manual Table of Contents I. Description...3 II. III. IV. Components...3 Storage...3 Features...3 V. General Composition of PCR Reaction
Phylogenetic Relationships among Worldwide Populations of the Brown Bear Ursus arctos Author(s): Tamako Matsuhashi, Ryuichi Masuda, Tsutomu Mano, Koichi Murata, and Awirmed Aiurzaniin Source: Zoological
Lit. # L0126 Rev. 8/99 Competitive PCR Guide Table of Contents I. What is Competitive PCR? A. Difficulties of Quantitative Analysis in Normal PCR... 2 B. Principle of Competitive PCR... 3 C. Competitive
Page 1 of 5 Page designed and maintained by Octavian Henegariu (Email: Tavi's Yale email or Tavi's Yahoo email). As I am currently pursuing a new junior faculty position, the Yale URL and email may change
Computational Biology Class Presentation Gene Trees in Species Trees Wayne P. Maddison Published in Systematic Biology, Vol. 46, No.3 (Sept 1997), pp. 523-536 Overview Gene Trees and Species Trees Processes
A Modified Wizard SV Genomic DNA Purification System Protocol to Purify Genomic DNA... A Modified Wizard SV Genomic DNA Purification System Protocol to Purify Genomic DNA from Shed Reptile Skin ABSTRACT
Likelihood Ratio Tests for Detecting Positive Selection and Application to Primate Lysozyme Evolution Ziheng Yang Department of Biology, University College, London An excess of nonsynonymous substitutions
Unit 7 Study Guide Section 8.7: Mutations KEY CONCEPT Mutations are changes in DNA that may or may not affect phenotype. VOCABULARY mutation point mutation frameshift mutation mutagen MAIN IDEA: Some mutations
PHYML Online: A Web Server for Fast Maximum Likelihood-Based Phylogenetic Inference Stephane Guindon, F. Le Thiec, Patrice Duroux, Olivier Gascuel To cite this version: Stephane Guindon, F. Le Thiec, Patrice
Muscular Dystrophy A Case Study of Positional Cloning Described by Benjamin Duchenne (1868) X-linked recessive disease causing severe muscular degeneration. 100 % penetrance X d Y affected male Frequency
Sanger Sequencing and Quality Assurance Zbigniew Rudzki Department of Pathology University of Melbourne Sanger DNA sequencing The era of DNA sequencing essentially started with the publication of the enzymatic
A branch-and-bound algorithm for the inference of ancestral amino-acid sequences when the replacement rate varies among sites Tal Pupko 1,*, Itsik Pe er 2, Masami Hasegawa 1, Dan Graur 3, and Nir Friedman
IMBB 2013 Genomic DNA purifica8on Why purify DNA? The purpose of DNA purifica8on from the cell/8ssue is to ensure it performs well in subsequent downstream applica8ons, e.g. Polymerase Chain Reac8on (PCR),
Name Period The AP Biology exam has reached into this chapter for essay questions on a regular basis over the past 15 years. Student responses show that biotechnology is a difficult topic. This chapter
POLYMERASE CHAIN REACTION (PCR) The polymerase chain reaction (PCR) is a technique used to amplify specific segments of DNA that may range in size from ca. 200-2000 or more base pairs. Two recent papers
Troubleshooting Sequencing Data Troubleshooting Sequencing Data No recognizable sequence (see page 7-10) Insufficient Quantitate the DNA. Increase the amount of DNA in the sequencing reactions. See page
Mitochondrial DNA Analysis Lineage Markers Lineage markers are passed down from generation to generation without changing Except for rare mutation events They can help determine the lineage (family tree)
Overview KAPAHiFi HotStart is the engineered KAPAHiFi DNA Polymerase with an antibody-based hot start technology and improved buffer system. KAPAHiFi HotStart exhibits: World leading fidelity confirmed
How many of you have checked out the web site on protein-dna interactions? Example of an approximately 40,000 probe spotted oligo microarray with enlarged inset to show detail. Find and be ready to discuss
How to Build a Phylogenetic Tree Phylogenetics tree is a structure in which species are arranged on branches that link them according to their relationship and/or evolutionary descent. A typical rooted
Genetics 10201232 Faculty of Agriculture and Veterinary Medicine Instructor: Dr. Jihad Abdallah Topic 15:Recombinant DNA Technology 1 Recombinant DNA Technology Recombinant DNA Technology is the use of
COMPARING DNA SEQUENCES TO DETERMINE EVOLUTIONARY RELATIONSHIPS AMONG MOLLUSKS OVERVIEW In the online activity Biodiversity and Evolutionary Trees: An Activity on Biological Classification, you generated
CHAPTER ONE: INTRODUCTION... 1 STUDY GOALS AND OBJECTIVES... 1 REASONS FOR DOING THIS STUDY... 1 SCOPE AND FORMAT... 1 METHODOLOGY AND INFORMATION SOURCES... 2 INTENDED AUDIENCE... 2 ANALYST CREDENTIALS...
Revised Fall 2011 The Techniques of Molecular Biology: Forensic DNA Fingerprinting The techniques of molecular biology are used to manipulate the structure and function of molecules such as DNA and proteins
BIOLOGY & EVOLUTION OF THE BIVALVIA Cambridge 14-17 September 1999 Mitochondrial and nuclear DNA phylogeography of two cupped oysters Crassostrea gigas and Crassostrea angulata P. Boudry & A. Huvet Laboratoire
Genetic Engineering Science as a Way of Knowing Q: How and why do scientists manipulate DNA in living cells? 15.1 How do humans take advantage of naturally occurring variation among organisms? WHAT I KNOW
Protocols Internal transcribed spacer region (ITS) region Niklaus J. Grünwald, Frank N. Martin, and Meg M. Larsen (2013) The nuclear ribosomal RNA (rrna) genes (small subunit, large subunit and 5.8S) are
Lectures 26 and 27 recombinant DNA technology I. oal of genetics A. historically - easy to isolate total DNA - difficult to isolate individual gene B. recombinant DNA technology C. why get the gene? 1.
Microbiology Laboratory: MOLECULAR IDENTIFICATION OF UNKNOWN BACTERIA Classical Microbiology courses are typically structured to introduce the identification of bacterial species using a series of biochemical
Lab 1: Who s Your Daddy? (AKA DNA Purification and PCR) Goals of the lab: 1. To understand how DNA s chemical properties can be exploited for purification 2. To learn practical applications of DNA purification
Supplementary Information - PCR amplification PCR amplification reactions for the partial mitochondrial cytochrome oxidase subunit I (COI), the ribosomal 16S rdna gene and a fragment of the nuclear single
Next Generation Polymerase Chain Reaction Developed by Nobel laureate Kary Mullis in the 1980s, Polymerase Chain Reaction (PCR) is a molecular technology that allows fast and in vitro. It has since become
User Manual Mir-X mirna First-Strand Synthesis Kit User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Clontech Laboratories, Inc.
TECH NOTE Characterization of CRISPR/Cas9 introduced Mutations using the Guide it Indel Identification Kit Streamlined method for characterizing the variety of indels introduced by genome editing technologies:
Promega Notes Number 71, 1999, p. 10 Blunt-End Pfu DNA Polymerase- Generated PCR Fragments into pgem -T Vector Systems By Kimberly Knoche, Ph.D., and Dan Kephart, Ph.D. Promega Corporation Corresponding
Transcription Animations Name: Lew Ports Biology Place http://www.lewport.wnyric.org/jwanamaker/animations/protein%20synthesis%20-%20long.html Protein is the making of proteins from the information found
Note: When any ambiguity of interpretation is found in this provisional translation, the Japanese text shall prevail. Chapter 2 Biological Inventions (Applied to any patent applications filed on or after
Chironomid DNA Barcode Database Search System User Manual National Institute for Environmental Studies Center for Environmental Biology and Ecosystem Studies December 2015 Contents 1. Overview 1 2. Search
Name Period Chapter 20: Biotechnology The AP Biology exam has reached into this chapter for essay questions on a regular basis over the past 15 years. Student responses show that biotechnology is a difficult
851 NOTE / NOTE Benchmarking DNA barcodes: an assessment using available primate sequences Mehrdad Hajibabaei, Gregory A.C. Singer, and Donal A. Hickey Abstract: DNA barcoding has been recently promoted
Lab 2/Phylogenetics/September 16, 2002 1 Read: Tudge Chapter 2 PHYLOGENETICS Objective of the Lab: To understand how DNA and protein sequence information can be used to make comparisons and assess evolutionary
DNA Fingerprinting Unless they are identical twins, individuals have unique DNA DNA fingerprinting The name used for the unambiguous identifying technique that takes advantage of differences in DNA sequence
Eötvös Lóránd University Biology Doctorate School Classical and molecular genetics program Project leader: Dr. László Orosz, corresponding member of HAS PHYLOGENY AND EVOLUTION OF NEWCASTLE DISEASE VIRUS
IFU022 v140202 Research Use Only Instructions For Use Part III Data Analysis for Ion Torrent Sequencing MANUFACTURER: Multiplicom N.V. Galileilaan 18 2845 Niel Belgium Revision date: August 21, 2014 Page
DNA, Replication and Transcription Bob Jesberg NSTA Conference Boston, MA April 3, 2014 1 Workshop Agenda Looking at DNA and Forensics The DNA, Replication i and Transcription i Set DNA Ladder The Double
SAM Teacher s Guide DNA to Proteins Note: Answers to activity and homework questions are only included in the Teacher Guides available after registering for the SAM activities, and not in this sample version.
Journal of Bioinformatics and Sequence Analysis Vol. 3(1), pp. 1-5, January 2011 Available online at http://www.academicjournals.org/jbsa ISSN 2141-2464 2011 Academic Journals Full length Research Paper
Protein Sequence Analysis - Overview - UDEL Workshop Raja Mazumder Research Associate Professor, Department of Biochemistry and Molecular Biology Georgetown University Medical Center Topics Why do protein
DNA and Forensic Science Micah A. Luftig * Stephen Richey ** I. INTRODUCTION This paper represents a discussion of the fundamental principles of DNA technology as it applies to forensic testing. A brief
Log In Sign Up Biotechnology Test Test 15 Matching Questions Regenerate Test 1. Plasmid 2. PCR Process 3. humulin 4. pluripotent 5. polymerase chain reaction (PCR) a b Is much smaller than the human genome,
Why? Genetic Mutations What mistakes can occur when DNA is replicated? The genes encoded in your DNA result in the production of proteins that perform specific functions within your cells. Various environmental
Supplementary Material for: Multiple Losses of Flight and Recent Speciation in Steamer Ducks Tara L. Fulton, Brandon Letts, and Beth Shapiro 1. Supplementary Tables Supplementary Table S1. Sample information.
Polymerase Chain Reaction (PCR) Paul C Winter, Belfast City Hospital, Belfast, UK The polymerase chain reaction is a technique that allows DNA molecules of interest (usually gene sequences) to be copied
05 PCR primer design 05.01 Design criteria PCR: an overview (from 02.01) PCR fidelity and optimization Primer design alessandro bogliolo isti information science and technology institute 1/20 Polymerase
Evolution at Two Levels in Humans and Chimpanzees Mary-Claire King and A.C. Wilson What did we know prior to 1975? 1700 s: Linnaeus and others of that time considered Great Apes to be the closest relatives
BIOTECHNOLOGY What can we do with DNA? Biotechnology Manipulation of biological organisms or their components for research and industrial purpose Usually manipulate DNA itself How to study individual gene?
Amazing DNA facts These facts can form the basis of a quiz (for example, how many base pairs are there in the human genome?). Students should be familiar with most of this material, so the quiz could be
Biotechnology and reporter genes Here, a lentivirus is used to carry foreign DNA into chickens. A reporter gene (GFP)indicates that foreign DNA has been successfully transferred. Recombinant DNA continued
Workshop Program Biodiversity, Butterflies & DNA Barcoding Wednesday 17 February 2016 10:00am - 15:00pm Computer Laboratory, Tunku Abdul Rahman University College, Malaysia The workshop is jointly organized
Forensic DNA Testing Terminology ABI 310 Genetic Analyzer a capillary electrophoresis instrument used by forensic DNA laboratories to separate short tandem repeat (STR) loci on the basis of their size.
Lecture 13: DNA Technology DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology DNA Sequencing determine order of nucleotides in a strand of DNA > bases = A,
Nucleic Acid Techniques in Bacterial Systematics Edited by Erko Stackebrandt Department of Microbiology University of Queensland St Lucia, Australia and Michael Goodfellow Department of Microbiology University
Guideline for the submission of DNA sequences and associated annotations within the framework of Directive 2001/18/EC and Regulation (EC) No 1829/2003 European Reference Laboratory for Genetically Modified
Biotechnology Chapter 20: Biotechnology: DNA Technology & Genomics The BIG Questions How can we use our knowledge of DNA to: o Diagnose disease or defect? o Cure disease or defect? o Change/improve organisms?
-1895A APPLICATION INFORMATION Automated Solutions AUTOMATED METHODS FOR PCR SETUP ON THE IOMEK FX Lisa Knapp, Jason Fawcett, and Laura Pajak, Ph.D. eckman Coulter, Inc. Introduction Since the development
Recombinant DNA & Genetic Engineering g Genetic Manipulation: Tools Kathleen Hill Associate Professor Department of Biology The University of Western Ontario Tools for Genetic Manipulation DNA, RNA, cdna
Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue Application Guide... 2 I. Description... 2 II. Product Components... 2 III. Materials to be Supplied
LabGenius The world s most advanced synthetic DNA libraries Technical design notes firstname.lastname@example.org V1.5 NOV 15 Introduction OUR APPROACH LabGenius is a gene synthesis company focussed on the design and manufacture
1. A recombinant DNA molecules is one that is a. produced through the process of crossing over that occurs in meiosis b. constructed from DNA from different sources c. constructed from novel combinations
For Research Use PrimeSTAR GXL DNA Polymerase Product Manual Table of Contents I. Description...3 II. Components...3 III. Storage...3 IV. Protocols...4 V. Optimization of Parameters...6 VI. Electrophoresis,
Bioinformatics Resources at a Glance A Note about FASTA Format There are MANY free bioinformatics tools available online. Bioinformaticists have developed a standard format for nucleotide and protein sequences
1/12 Dideoxy DNA Sequencing Dideoxy DNA sequencing utilizes two steps: PCR (polymerase chain reaction) amplification of DNA using dideoxy nucleoside triphosphates (Figures 1 and 2)and denaturing polyacrylamide
SR3_3E S h i m a d z u R e v i e w DNA Analytical Technology to Identify the Actual Composition of Foods - Application of the MultiNA Microchip Electrophoresis System - by Yuji SOGABE Abstract Recently,
The Integrated Web Portal (IWP) Author(s): Rajan Man Bajracharya, Paribesh Pradhan, Bikash Dangol, Kiran Shakya, and Basanta Shrestha Source:, 30(2):178-181. Published By: International Mountain Society
Vector NTI Advance 11 Quick Start Guide Catalog no. 12605050, 12605099, 12605103 Version 11.0 December 15, 2008 12605022 Published by: Invitrogen Corporation 5791 Van Allen Way Carlsbad, CA 92008 U.S.A.
DnaSP, DNA polymorphism analyses by the coalescent and other methods. Author affiliation: Julio Rozas 1, *, Juan C. Sánchez-DelBarrio 2,3, Xavier Messeguer 2 and Ricardo Rozas 1 1 Departament de Genètica,