Players. Processes. Molecular Biology Recombinant DNA technology (So... you want to be a genetic engineer)

Size: px
Start display at page:

Download "Players. Processes. Molecular Biology Recombinant DNA technology (So... you want to be a genetic engineer)"

Transcription

1 lay Davis, 8/11/12 Molecular Biology Recombinant DN technology (So... you want to be a genetic engineer) Introduction, or, games to play with DN You have learned something of the structure of DN, its replication, and how the information is read by the cell to produce a functional molecule, often a protein. In this module we use the fluorescent green protein (F) that is made in a jellyfish as a starting point for exploring the techniques and tools of genetic engineering. The basic challenge is to transfer the genetic information from the jellyfish to bacteria such that the bacteria are fluorescent. In doing so we will pick up the basic principles and tools of genetic engineering and review the basic molecular biology of prokaryotes and eukaryotes. Reading Lehninger, 4th ed, pgs The ell, 3rd ed., pgs Need to know and understand: rocesses DN purification layers phenol extraction Transformation chemical (al 2 ) electroporation loning Recombinant DN enomic DN library Expression library Vectors - These are specialized pieces of DN that can carry, replicate and transfer pieces of DN that we insert in their respective host organisms. Typically in bacterial hosts: lasmid, osmid, hage λ, Bacterial rtificial hromosomes (Bs). lasmids, cosmids and BS all contain an ori, antibiotic resistance gene and a cloning site (MS). Restriction endonucleases T4 DN ligase 1

2 cdn DN Sequencing Reverse transcriptase E. coli DN ligase RNase H DN polymerase dnts rimer Template dnts ddnts DN polymerase el electrophoresis: Forward force on DN depends on charge of DN charge on DN depends on length mass of DN depends on length frictional force on DN depends on length of DN, and on conformation of DN, and on concentration of matrix log 10 (size) = -K 1( d) K 2 where: size = size of DN fragment in base pairs or kilobasebpairs d = migration distance (mobility) from start (origin) olymerase hain Reaction (R) rimer Template dnts thermostable DN polymerase thermal cycler olony hybridization (screening) preparing probe nitrocellulose replica denaturing hybridizing Hybridization Kinetics (This is the basis for southern and Northern blots and in situ hybridization) 100 o T( o ) <T m ssdn ssdn -> dsdn t The toolbox Enzymes - The enzymes and their activities listed on the following pages are our basic tools of molecular biology. In general each tool will perform a specific action on the target molecule, often DN or RN. Refer to this list as we go through procedures, collect and remember the activities as we go though the lessons. You do not need to remember them all, just the ones we use. 2

3 Enzyme activities: 1. ->3' polymerase activity T α γ β α γ β 2. 3' -> exonuclease activity T T 3.endonuclease activity 4. ->3' exonuclease activity 5. phosphatase activity 3

4 6. kinase activity γ β α γ γ β α 7. Ligase activity 8. ->3' RN polymerase activity U U 4

5 Enzymes E. coli DN polymerase I holoenzyme (ol I) activity: -3' DN polymerase substrate: primed DN activity:3'-> exonuclease substrate: dsdn, ssdn note: slower than polymerase activity activity: -3' exonuclease substrate: dsdn, ssdn note: very slow reaction on ds DN T4 DN polymerase or Klenow fragment of ol I activity: -3' DN polymerase substrate: primed DN S1 nuclease activity: endonuclease template: ssdn or nicked dsdn note: very active, since dsdn temporarily loses some base pairing due to thermal energy, S1 nuclease will eventually degrade dsdn. RNse activity: endonuclease substrate: ssrn RNase H activity: endonuclease, exonuclease substrate: dsrn or RN of DN/RN hybrid activity:3'-> exonuclease substrate: dsdn, ssdn T4 DN ligase: activity: Ligase substrate: dsdn with blunt or cohesive ends or nicked dsdn. E. coli ligase activity: Ligase substrate: Double stranded DN with cohesive ends or nicked dsdn Reverse transcriptase activity: -3' DN polymerase template: primed DN or RN DNase I activity: endonuclease substrate:dsdn or ssdn T7 or T3 or S6 polymerases activity: ->3' RN polymerase substrate: dsdn with T7 or T3 or S6 promoter sequence Restriction endonucleases activity: double stranded endonuclease substrate: dsdn with recognition sequence...nntnn... eg:...nntnn... BamHI T4 polynucleotide kinase activity: kinase template: of DN or RN...NN...NNT NN... TNN... Bacterial lkaline phosphatase (B) alf-intestinal phosphatase (I) activity: phosphatase template: phosphate of DN or RN (ss or ds). 5

6 The Story of a rotein, its Travels, and How it hanged on the Way. The reen Fluorescent rotein (F) is made by a jellyfish. Two views of the structure of F as determined by X-ray crystallography. The gold arrows are b-sheets. 6

7 The information to make F can be moved to other organisms. E. coli Fluorescent E. coli colonies on a petri dish Fluorescent E. coli under the microscope If you play with the sequence of the F protein, you can change is color. 7

8 The F sequence can be added to other eukaryotic cells and animals 8

Molecular Cell Biology - Problem Drill 11: Recombinant DNA

Molecular Cell Biology - Problem Drill 11: Recombinant DNA Molecular Cell Biology - Problem Drill 11: Recombinant DNA Question No. 1 of 10 1. Which of the following statements about the sources of DNA used for molecular cloning is correct? Question #1 (A) cdna

More information

Computational Biology I LSM5191

Computational Biology I LSM5191 Computational Biology I LSM5191 Lecture 5 Notes: Genetic manipulation & Molecular Biology techniques Broad Overview of: Enzymatic tools in Molecular Biology Gel electrophoresis Restriction mapping DNA

More information

Lecture Four. Molecular Approaches I: Nucleic Acids

Lecture Four. Molecular Approaches I: Nucleic Acids Lecture Four. Molecular Approaches I: Nucleic Acids I. Recombinant DNA and Gene Cloning Recombinant DNA is DNA that has been created artificially. DNA from two or more sources is incorporated into a single

More information

Gene Cloning & DNA Analysis

Gene Cloning & DNA Analysis CSS451 CSS/HRT 451 Gene Cloning & DNA Analysis Chapter 4-5 T-DNA LB auxin cytokin opine Oncogenic genes RB vir genes ori opine catabolism Guo-qing Song Part 1 Basic principles Gene Cloning & DNA Analysis

More information

M Keramatipour 2. M Keramatipour 1. M Keramatipour 4. M Keramatipour 3. M Keramatipour 5. M Keramatipour

M Keramatipour 2. M Keramatipour 1. M Keramatipour 4. M Keramatipour 3. M Keramatipour 5. M Keramatipour Molecular Cloning Methods Mohammad Keramatipour MD, PhD keramatipour@tums.ac.ir Outline DNA recombinant technology DNA cloning co Cell based PCR PCR-based Some application of DNA cloning Genomic libraries

More information

2054, Chap. 14, page 1

2054, Chap. 14, page 1 2054, Chap. 14, page 1 I. Recombinant DNA technology (Chapter 14) A. recombinant DNA technology = collection of methods used to perform genetic engineering 1. genetic engineering = deliberate modification

More information

CHAPTER 9 DNA Technologies

CHAPTER 9 DNA Technologies CHAPTER 9 DNA Technologies Recombinant DNA Artificially created DNA that combines sequences that do not occur together in the nature Basis of much of the modern molecular biology Molecular cloning of genes

More information

Lecture 25 (11/15/17)

Lecture 25 (11/15/17) Lecture 25 (11/15/17) Reading: Ch9; 328-332 Ch25; 990-995, 1005-1012 Problems: Ch9 (study-guide: applying); 1,2 Ch9 (study-guide: facts); 7,8 Ch25 (text); 1-3,5-7,9,10,13-15 Ch25 (study-guide: applying);

More information

AP Biology. Chapter 20. Biotechnology: DNA Technology & Genomics. Biotechnology. The BIG Questions. Evolution & breeding of food plants

AP Biology. Chapter 20. Biotechnology: DNA Technology & Genomics. Biotechnology. The BIG Questions. Evolution & breeding of food plants What do you notice about these phrases? radar racecar Madam I m Adam Able was I ere I saw Elba a man, a plan, a canal, Panama Was it a bar or a bat I saw? Chapter 20. Biotechnology: DNA Technology & enomics

More information

DNA Technology. Asilomar Singer, Zinder, Brenner, Berg

DNA Technology. Asilomar Singer, Zinder, Brenner, Berg DNA Technology Asilomar 1973. Singer, Zinder, Brenner, Berg DNA Technology The following are some of the most important molecular methods we will be using in this course. They will be used, among other

More information

Manipulation of Purified DNA

Manipulation of Purified DNA Manipulation of Purified DNA To produce the recombinant DNA molecule, the vector, as well as the DNA to be cloned, must be cut at specific points and then joined together in a controlled manner by DNA

More information

Recombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology.

Recombinant DNA Technology. The Role of Recombinant DNA Technology in Biotechnology. yeast. Biotechnology. Recombinant DNA technology. PowerPoint Lecture Presentations prepared by Mindy Miller-Kittrell, North Carolina State University C H A P T E R 8 Recombinant DNA Technology The Role of Recombinant DNA Technology in Biotechnology Biotechnology?

More information

CHAPTER 20 DNA TECHNOLOGY AND GENOMICS. Section A: DNA Cloning

CHAPTER 20 DNA TECHNOLOGY AND GENOMICS. Section A: DNA Cloning Section A: DNA Cloning 1. DNA technology makes it possible to clone genes for basic research and commercial applications: an overview 2. Restriction enzymes are used to make recombinant DNA 3. Genes can

More information

Learning Objectives :

Learning Objectives : Learning Objectives : Understand the basic differences between genomic and cdna libraries Understand how genomic libraries are constructed Understand the purpose for having overlapping DNA fragments in

More information

Multiple choice questions (numbers in brackets indicate the number of correct answers)

Multiple choice questions (numbers in brackets indicate the number of correct answers) 1 Multiple choice questions (numbers in brackets indicate the number of correct answers) February 1, 2013 1. Ribose is found in Nucleic acids Proteins Lipids RNA DNA (2) 2. Most RNA in cells is transfer

More information

Genetic Engineering & Recombinant DNA

Genetic Engineering & Recombinant DNA Genetic Engineering & Recombinant DNA Chapter 10 Copyright The McGraw-Hill Companies, Inc) Permission required for reproduction or display. Applications of Genetic Engineering Basic science vs. Applied

More information

Cyclin- Dependent Protein Kinase. Function

Cyclin- Dependent Protein Kinase. Function Complex Cyclin Cyclin- Dependent rotein Kinase Function promotes passage through restriction G1-CDK Complex Cyclin D CDK4 or CDK6 point in late G1 G1/S-CDK Complex Cyclin E CDK2 commits the cell to DA

More information

Genetics Lecture 21 Recombinant DNA

Genetics Lecture 21 Recombinant DNA Genetics Lecture 21 Recombinant DNA Recombinant DNA In 1971, a paper published by Kathleen Danna and Daniel Nathans marked the beginning of the recombinant DNA era. The paper described the isolation of

More information

AP Biology Gene Expression/Biotechnology REVIEW

AP Biology Gene Expression/Biotechnology REVIEW AP Biology Gene Expression/Biotechnology REVIEW Multiple Choice Identify the choice that best completes the statement or answers the question. 1. Gene expression can be a. regulated before transcription.

More information

Bacterial DNA replication

Bacterial DNA replication Bacterial DNA replication Summary: What problems do these proteins solve? Tyr OH attacks PO4 and forms a covalent intermediate Structural changes in the protein open the gap by 20 Å! 1 Summary: What problems

More information

CHAPTER 08: RECOMBINANT DNA TECHNOLOGY Pearson Education, Inc.

CHAPTER 08: RECOMBINANT DNA TECHNOLOGY Pearson Education, Inc. CHAPTER 08: RECOMBINANT DNA TECHNOLOGY The Role of Recombinant DNA Technology in Biotechnology Biotechnology the use of microorganisms to make practical products Recombinant DNA technology Intentionally

More information

MCB 150: The Molecular and Cellular Basis of Life

MCB 150: The Molecular and Cellular Basis of Life MCB 150 The Molecular and Cellular Basis of Life Plasmids and Genetic Engineering I Today s Learning Catalytics Session ID is: 20000966 1 Announcements: Check gradebook for discrepancies by Wednesday at

More information

Molecular Biology, Lecture 3 DNA Replication

Molecular Biology, Lecture 3 DNA Replication Molecular Biology, Lecture 3 DNA Replication We will continue talking about DNA replication. We have previously t discussed the structure of DNA. DNA replication is the copying of the whole DNA content

More information

BIOTECHNOLOGY OLD BIOTECHNOLOGY (TRADITIONAL BIOTECHNOLOGY) MODERN BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY.

BIOTECHNOLOGY OLD BIOTECHNOLOGY (TRADITIONAL BIOTECHNOLOGY) MODERN BIOTECHNOLOGY RECOMBINANT DNA TECHNOLOGY. BIOTECHNOLOGY Biotechnology can be defined as the use of micro-organisms, plant or animal cells or their components or enzymes from organisms to produce products and processes (services) useful to human

More information

BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA.

BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. Lab#2 BCH 462 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. Outlines: 1-Insertion of foreign gene to the plasmid. 2-Competent cell. 3-Transformation of bacterial cell.

More information

Chapter 20: Biotechnology

Chapter 20: Biotechnology 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

More information

1. A brief overview of sequencing biochemistry

1. A brief overview of sequencing biochemistry Supplementary reading materials on Genome sequencing (optional) The materials are from Mark Blaxter s lecture notes on Sequencing strategies and Primary Analysis 1. A brief overview of sequencing biochemistry

More information

Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, ; ; 330 PCR, ; 329.

Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, ; ; 330 PCR, ; 329. Lecture 3 (FW) January 28, 2009 Cloning of DNA; PCR amplification Reading assignment: Cloning, 240-245; 286-87; 330 PCR, 270-274; 329. Take Home Lesson(s) from Lecture 2: 1. DNA is a double helix of complementary

More information

DNA Technology. B. Using Bacteria to Clone Genes: Overview:

DNA Technology. B. Using Bacteria to Clone Genes: Overview: DNA Technology A. Basic Vocabulary: is DNA from 2 different sources that is combined. is the direct manipulation of genes for practical purposes. literally means or in a test tube or flask. is the manipulation

More information

SEQUENCING DNA. Jos. J. Schall Biology Department University of Vermont

SEQUENCING DNA. Jos. J. Schall Biology Department University of Vermont SEQUENCING DNA Jos. J. Schall Biology Department University of Vermont SEQUENCING DNA Start with PCR product (your end result of a PCR). Remember, your template DNA in the PCR was extracted DNA that included

More information

Synthetic Biology for

Synthetic Biology for Synthetic Biology for Plasmids and DNA Digestion Plasmids Plasmids are small DNA molecules that are separate from chromosomal DNA They are most commonly found as double stranded, circular DNA Typical plasmids

More information

NAME TA SEC Problem Set 3 FRIDAY March 5, Problem sets will NOT be accepted late.

NAME TA SEC Problem Set 3 FRIDAY March 5, Problem sets will NOT be accepted late. MIT Department of Biology 7.013: Introductory Biology - Spring 2004 Instructors: Professor Hazel Sive, Professor Tyler Jacks, Dr. laudette ardel NME T SE 7.013 Problem Set 3 FRIDY March 5, 2004 Problem

More information

2 Gene Technologies in Our Lives

2 Gene Technologies in Our Lives CHAPTER 15 2 Gene Technologies in Our Lives SECTION Gene Technologies and Human Applications KEY IDEAS As you read this section, keep these questions in mind: For what purposes are genes and proteins manipulated?

More information

Recombinant DNA Technology

Recombinant DNA Technology History of recombinant DNA technology Recombinant DNA Technology (DNA cloning) Majid Mojarrad Recombinant DNA technology is one of the recent advances in biotechnology, which was developed by two scientists

More information

Covalently bonded sugar-phosphate backbone with relatively strong bonds keeps the nucleotides in the backbone connected in the correct sequence.

Covalently bonded sugar-phosphate backbone with relatively strong bonds keeps the nucleotides in the backbone connected in the correct sequence. Unit 14: DNA Replication Study Guide U7.1.1: DNA structure suggested a mechanism for DNA replication (Oxford Biology Course Companion page 347). 1. Outline the features of DNA structure that suggested

More information

Genetic Engineering Genetic engineering primarily involves the manipulation of genetic material (DNA) to achieve the desired goal in a pre-determined

Genetic Engineering Genetic engineering primarily involves the manipulation of genetic material (DNA) to achieve the desired goal in a pre-determined Genetic Engineering Genetic engineering primarily involves the manipulation of genetic material (DNA) to achieve the desired goal in a pre-determined way. Some other terms are also in common use to describe

More information

DNA REPLICATION. Anna Onofri Liceo «I.Versari»

DNA REPLICATION. Anna Onofri Liceo «I.Versari» DNA REPLICATION Anna Onofri Liceo «I.Versari» Learning objectives 1. Understand the basic rules governing DNA replication 2. Understand the function of key proteins involved in a generalised replication

More information

Green Fluorescent Protein (GFP) Purification. Hydrophobic Interaction Chromatography

Green Fluorescent Protein (GFP) Purification. Hydrophobic Interaction Chromatography Green Fluorescent Protein (GFP) Purification Hydrophobic Interaction Chromatography What is the GFP gene? GFP is a green fluorescent protein that is normally found in jellyfish. It has been engineered

More information

How Do You Clone a Gene?

How Do You Clone a Gene? S-20 Edvo-Kit #S-20 How Do You Clone a Gene? Experiment Objective: The objective of this experiment is to gain an understanding of the structure of DNA, a genetically engineered clone, and how genes are

More information

The Molecul Chapter ar Basis 16: The M of olecular Inheritance Basis of Inheritance Fig. 16-1

The Molecul Chapter ar Basis 16: The M of olecular Inheritance Basis of Inheritance Fig. 16-1 he Chapter Molecular 16: he Basis Molecular of Inheritance Basis of Inheritance Fig. 16-1 dditional Evidence hat DN Is the Genetic Material It was known that DN is a polymer of nucleotides, each consisting

More information

The replication of DNA Kornberg 1957 Meselson and Stahl 1958 Cairns 1963 Okazaki 1968 DNA Replication The driving force for DNA synthesis. The addition of a nucleotide to a growing polynucleotide

More information

Biotechnology and Genomics in Public Health. Sharon S. Krag, PhD Johns Hopkins University

Biotechnology and Genomics in Public Health. Sharon S. Krag, PhD Johns Hopkins University This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike License. Your use of this material constitutes acceptance of that license and the conditions of use of materials on this

More information

Storage and Expression of Genetic Information

Storage and Expression of Genetic Information Storage and Expression of Genetic Information 29. DNA structure, Replication and Repair ->Ch 25. DNA metabolism 30. RNA Structure, Synthesis and Processing ->Ch 26. RNA metabolism 31. Protein Synthesis

More information

MOLECULAR BASIS OF INHERITANCE

MOLECULAR BASIS OF INHERITANCE MOLECULAR BASIS OF INHERITANCE C H A P T E R 1 6 as genetic material? Deducted that is the genetic material Initially worked by studying bacteria & the viruses that infected them 1928 Frederick Griffiths

More information

CHAPTER 16 MOLECULAR BASIS OF INHERITANCE

CHAPTER 16 MOLECULAR BASIS OF INHERITANCE CHAPTER 16 MOLECULAR BASIS OF INHERITANCE DNA as genetic material? Deducted that DNA is the genetic material Initially worked by studying bacteria & the viruses that infected them 1928 Frederick Griffiths

More information

INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist

INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist INTRODUCTION TO REVERSE TRANSCRIPTION PCR (RT-PCR) ABCF 2016 BecA-ILRI Hub, Nairobi 21 st September 2016 Roger Pelle Principal Scientist Objective of PCR To provide a solution to one of the most pressing

More information

DNA REPLICATION & REPAIR

DNA REPLICATION & REPAIR DNA REPLICATION & REPAIR Table of contents 1. DNA Replication Model 2. DNA Replication Mechanism 3. DNA Repair: Proofreading 1. DNA Replication Model Replication in the cell cycle 3 models of DNA replication

More information

DNA Replication II Biochemistry 302. January 25, 2006

DNA Replication II Biochemistry 302. January 25, 2006 DNA Replication II Biochemistry 302 January 25, 2006 Following in Dad s footsteps Original A. Kornberg E. coli DNA Pol I is a lousy replicative enzyme. 400 molecules/cell but ~2 replication forks/cell

More information

BIOTECHNOLOGY : PRINCIPLES AND PROCESSES Biotechnologydeals with techniques of using live organisms or enzymes from organisms to produce products and

BIOTECHNOLOGY : PRINCIPLES AND PROCESSES Biotechnologydeals with techniques of using live organisms or enzymes from organisms to produce products and BIOTECHNOLOGY : PRINCIPLES AND PROCESSES Biotechnologydeals with techniques of using live organisms or enzymes from organisms to produce products and processes useful to humans. Traditional form based

More information

DNA Replication I Biochemistry 302. Bob Kelm January 24, 2005

DNA Replication I Biochemistry 302. Bob Kelm January 24, 2005 DNA Replication I Biochemistry 302 Bob Kelm January 24, 2005 Watson Crick prediction: Each stand of parent DNA serves as a template for synthesis of a new complementary daughter strand Fig. 4.12 Proof

More information

Amira A. AL-Hosary PhD of infectious diseases Department of Animal Medicine (Infectious Diseases) Faculty of Veterinary Medicine Assiut

Amira A. AL-Hosary PhD of infectious diseases Department of Animal Medicine (Infectious Diseases) Faculty of Veterinary Medicine Assiut Amira A. AL-Hosary PhD of infectious diseases Department of Animal Medicine (Infectious Diseases) Faculty of Veterinary Medicine Assiut University-Egypt Restriction Endonucleases, (cutting dna) (ligation)

More information

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity

The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity Promega Notes Magazine Number 62, 1997, p. 02 The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity By Christine Andrews and Scott Lesley Promega

More information

Bio 101 Sample questions: Chapter 10

Bio 101 Sample questions: Chapter 10 Bio 101 Sample questions: Chapter 10 1. Which of the following is NOT needed for DNA replication? A. nucleotides B. ribosomes C. Enzymes (like polymerases) D. DNA E. all of the above are needed 2 The information

More information

HiPer Plasmid DNA Cloning Teaching Kit

HiPer Plasmid DNA Cloning Teaching Kit HiPer Plasmid DNA Cloning Teaching Kit Product Code: HTBM022 Number of experiments that can be performed: 5 Duration of Experiment: 4 days Day 1- Preparation of media and revival of E. coli Host Day2-

More information

Lecture 24: DNA replication. Lecture Outline. Chromosomes. Review

Lecture 24: DNA replication. Lecture Outline. Chromosomes. Review Lecture 4: D replication Lecture Outline Figure 6.7a, c nm (c) Space-filling model Overview of D replication D has polarity (5, 3 O) Rules of Replication Details of the replication machine roofreading

More information

Molecular Biology: DNA sequencing

Molecular Biology: DNA sequencing Molecular Biology: DNA sequencing Author: Prof Marinda Oosthuizen Licensed under a Creative Commons Attribution license. SEQUENCING OF LARGE TEMPLATES As we have seen, we can obtain up to 800 nucleotides

More information

3 Designing Primers for Site-Directed Mutagenesis

3 Designing Primers for Site-Directed Mutagenesis 3 Designing Primers for Site-Directed Mutagenesis 3.1 Learning Objectives During the next two labs you will learn the basics of site-directed mutagenesis: you will design primers for the mutants you designed

More information

Manipulating DNA. Nucleic acids are chemically different from other macromolecules such as proteins and carbohydrates.

Manipulating DNA. Nucleic acids are chemically different from other macromolecules such as proteins and carbohydrates. Lesson Overview 14.3 Studying the Human Genome Nucleic acids are chemically different from other macromolecules such as proteins and carbohydrates. Nucleic acids are chemically different from other macromolecules

More information

Genetic Fingerprinting

Genetic Fingerprinting Genetic Fingerprinting Introduction DA fingerprinting In the R & D sector: -involved mostly in helping to identify inherited disorders. In forensics: -identification of possible suspects involved in offences.

More information

BCMB Chapters 34 & 35 DNA Replication and Repair

BCMB Chapters 34 & 35 DNA Replication and Repair BCMB 3100 - Chapters 34 & 35 DNA Replication and Repair Semi-conservative DNA replication DNA polymerase DNA replication Replication fork; Okazaki fragments Sanger method for DNA sequencing DNA repair

More information

Gene Expression Technology

Gene Expression Technology Gene Expression Technology Bing Zhang Department of Biomedical Informatics Vanderbilt University bing.zhang@vanderbilt.edu Gene expression Gene expression is the process by which information from a gene

More information

DNA Structure and Analysis. Chapter 4: Background

DNA Structure and Analysis. Chapter 4: Background DNA Structure and Analysis Chapter 4: Background Molecular Biology Three main disciplines of biotechnology Biochemistry Genetics Molecular Biology # Biotechnology: A Laboratory Skills Course explorer.bio-rad.com

More information

Some types of Mutagenesis

Some types of Mutagenesis Mutagenesis What Is a Mutation? Genetic information is encoded by the sequence of the nucleotide bases in DNA of the gene. The four nucleotides are: adenine (A), thymine (T), guanine (G), and cytosine

More information

Gene Transfer 11/4/13. Fredrick Griffith in the 1920s did an experiment. Not until 1944 was DNA shown to be the moveable element

Gene Transfer 11/4/13. Fredrick Griffith in the 1920s did an experiment. Not until 1944 was DNA shown to be the moveable element Gene Transfer Fredrick Griffith in the 1920s did an experiment. Not until 19 was DN shown to be the moveable element Dead pathogen cells able to make a capsule were able to pass this ability to the live

More information

Chapter 8 Recombinant DNA Technology. 10/1/ MDufilho

Chapter 8 Recombinant DNA Technology. 10/1/ MDufilho Chapter 8 Recombinant DNA Technology 10/1/2017 1 MDufilho The Role of Recombinant DNA Technology in Biotechnology Biotechnology? Recombinant deoxyribonucleic acid (DNA) technology Intentionally modifying

More information

Directe d Mutagenesis

Directe d Mutagenesis Directe d Mutagenesis A Practical Approac h M. J. McPHERSON 1. Mutagenesis facilitated by the removal or introduction of unique restriction sites 1 P. Carte r 1. Introduction to site-directed mutagenesis

More information

Cloning Vectors Ameer Effat M. Elfarash

Cloning Vectors Ameer Effat M. Elfarash Cloning Vectors Ameer Effat M. Elfarash Dept. of Genetics Fac. of Agriculture, Assiut Univ. amir_effat@yahoo.com CLONING VECTORS Cloning vectors are DNA molecules that are used to "transport" cloned sequences

More information

Chapter 15 Gene Technologies and Human Applications

Chapter 15 Gene Technologies and Human Applications Chapter Outline Chapter 15 Gene Technologies and Human Applications Section 1: The Human Genome KEY IDEAS > Why is the Human Genome Project so important? > How do genomics and gene technologies affect

More information

MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien

MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien Introduction MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien The field of molecular genetics has resulted in a number of practical applications that have been of tremendous

More information

Molecular Biology: DNA, gene, chromosome and genome (Outline)

Molecular Biology: DNA, gene, chromosome and genome (Outline) Molecular Biology: D, gene, chromosome and genome (utline) ucleic acid structure and composition D and R Base-pairing rule in D Definition of D, gene, chromosome and genome. D structure and chemical bonds

More information

BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005

BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005 BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005 Lab Exercise 7 Drosophila crosses, three weeks Vocabulary: phenotype, genotype, gene, allele, locus (loci), sex chromosomes, autosomes, homozygous,

More information

Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami

Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami Reverse transcription-pcr (rt-pcr) Dr. Hani Alhadrami hanialhadrami@kau.edu.sa www.hanialhadrami.kau.edu.sa Overview Several techniques are available to detect and analyse RNA. Examples of these techniques

More information

Cloning in bacteria. Presenter: Vito Baraka (BSc,MSc Cand.)

Cloning in bacteria. Presenter: Vito Baraka (BSc,MSc Cand.) Cloning in bacteria Presenter: Vito Baraka (BSc,MSc Cand.) Introduction DNA cloning involves separating a specific gene or DNA segment from a larger chromosome, attaching it to a small molecule of carrier

More information

Cat # Box1 Box2. DH5a Competent E. coli cells CCK-20 (20 rxns) 40 µl 40 µl 50 µl x 20 tubes. Choo-Choo Cloning TM Enzyme Mix

Cat # Box1 Box2. DH5a Competent E. coli cells CCK-20 (20 rxns) 40 µl 40 µl 50 µl x 20 tubes. Choo-Choo Cloning TM Enzyme Mix Molecular Cloning Laboratories User Manual Version 3.3 Product name: Choo-Choo Cloning Kits Cat #: CCK-10, CCK-20, CCK-096, CCK-384 Description: Choo-Choo Cloning is a highly efficient directional PCR

More information

NOTES - CH 15 (and 14.3): DNA Technology ( Biotech )

NOTES - CH 15 (and 14.3): DNA Technology ( Biotech ) NOTES - CH 15 (and 14.3): DNA Technology ( Biotech ) Vocabulary Genetic Engineering Gene Recombinant DNA Transgenic Restriction Enzymes Vectors Plasmids Cloning Key Concepts What is genetic engineering?

More information

DNA replication. - proteins for initiation of replication; - proteins for polymerization of nucleotides.

DNA replication. - proteins for initiation of replication; - proteins for polymerization of nucleotides. DNA replication Replication represents the duplication of the genetic information encoded in DNA that is the crucial step in the reproduction of living organisms and the growth of multicellular organisms.

More information

Microbiology 微生物学 Spring-Summer

Microbiology 微生物学 Spring-Summer Microbiology 微生物学 2017 Spring-Summer Relevant Information and Resources Course slides can be found at http://mypage.zju.edu.cn/haichun 教学工作 Course-related questions will be answered through emails. Textbook:

More information

Cut Smarter with NEB Restriction Enzymes

Cut Smarter with NEB Restriction Enzymes ut Smarter with EB Restriction Enzymes A Smarter Look Keep track of your enzyme data with our new, streamlined protocol cards. These collectible cards contain all the information you need for setting up

More information

Replication. Obaidur Rahman

Replication. Obaidur Rahman Replication Obaidur Rahman DIRCTION OF DNA SYNTHESIS How many reactions can a DNA polymerase catalyze? So how many reactions can it catalyze? So 4 is one answer, right, 1 for each nucleotide. But what

More information

Test Prep Pretest. in the. the. whereas prokaryotic DNA contains only replication forks during replication. Skills Worksheet

Test Prep Pretest. in the. the. whereas prokaryotic DNA contains only replication forks during replication. Skills Worksheet Skills Worksheet Test Prep Pretest Complete each statement by writing the correct term or phrase in the space provided. 1. In 1928, Frederick Griffith found that the capsule that enclosed one strain of

More information

Enzymatic assembly of DNA molecules up to several hundred kilobases

Enzymatic assembly of DNA molecules up to several hundred kilobases nature methods Enzymatic assembly of DNA molecules up to several hundred kilobases Daniel G Gibson, Lei Young, Ray-Yuan Chuang, J Craig Venter, Clyde A Hutchison III & Hamilton O Smith Supplementary figures

More information

Chapter 20: Biotechnology

Chapter 20: Biotechnology Chapter 20: Biotechnology 1. DNA Sequencing 2. DNA Cloning 3. Studying Gene Expression 4. Manipulating Genomes 5. herapeutic & Diagnostic echniques 1. DNA Sequencing Chapter Reading pp. 409-412 DNA Sequencing

More information

Methods of Biomaterials Testing Lesson 3-5. Biochemical Methods - Molecular Biology -

Methods of Biomaterials Testing Lesson 3-5. Biochemical Methods - Molecular Biology - Methods of Biomaterials Testing Lesson 3-5 Biochemical Methods - Molecular Biology - Chromosomes in the Cell Nucleus DNA in the Chromosome Deoxyribonucleic Acid (DNA) DNA has double-helix structure The

More information

Genomics and Gene Recognition Genes and Blue Genes

Genomics and Gene Recognition Genes and Blue Genes Genomics and Gene Recognition Genes and Blue Genes November 1, 2004 Prokaryotic Gene Structure prokaryotes are simplest free-living organisms studying prokaryotes can give us a sense what is the minimum

More information

DNA replication: Enzymes link the aligned nucleotides by phosphodiester bonds to form a continuous strand.

DNA replication: Enzymes link the aligned nucleotides by phosphodiester bonds to form a continuous strand. DNA replication: Copying genetic information for transmission to the next generation Occurs in S phase of cell cycle Process of DNA duplicating itself Begins with the unwinding of the double helix to expose

More information

Sample Question Paper Session Class XII Biotechnology (045) Marking Scheme

Sample Question Paper Session Class XII Biotechnology (045) Marking Scheme Sample Question Paper Session 2015-16 Class XII Biotechnology (045) Marking Scheme SECTION-A 1. Methylation Student will add a methyl group to one or two bases within the sequence recognized by enzyme.

More information

American Society of Cytopathology Core Curriculum in Molecular Biology

American Society of Cytopathology Core Curriculum in Molecular Biology American Society of Cytopathology Core Curriculum in Molecular Biology American Society of Cytopathology Core Curriculum in Molecular Biology Chapter 3 Molecular Techniques Alternatives to PCR, Part I

More information

Chapter 18 Review Page 1

Chapter 18 Review Page 1 hapter 18 Review Page 1 1 In this diagram of a virus, the pointer is indicating the virus's capsid. genome. envelope. mitochondria. microfilaments. 2 Viral N makes mrn by the process of lysis. infection.

More information

NUCLEIC ACIDS Genetic material of all known organisms DNA: deoxyribonucleic acid RNA: ribonucleic acid (e.g., some viruses)

NUCLEIC ACIDS Genetic material of all known organisms DNA: deoxyribonucleic acid RNA: ribonucleic acid (e.g., some viruses) NUCLEIC ACIDS Genetic material of all known organisms DNA: deoxyribonucleic acid RNA: ribonucleic acid (e.g., some viruses) Consist of chemically linked sequences of nucleotides Nitrogenous base Pentose-

More information

STRUCTURE AND DIAGNOSTIC APPLICATIONS OF DNA

STRUCTURE AND DIAGNOSTIC APPLICATIONS OF DNA STRUCTURE AND DIAGNOSTIC APPLICATIONS OF DNA UNIVERSITY OF PAPUAN NEW GUINEA SCHOOL OF MEDICINE AND HEALTH SCIENCES DIVISION OF BASIC MEDICAL SCIENCES DISCIPLINE OF BIOCHEMISTRY & MOLECULAR BIOLOGY LECTURE

More information

Biochemistry. DNA Polymerase. Structure and Function of Biomolecules II. Principal Investigator

Biochemistry. DNA Polymerase. Structure and Function of Biomolecules II. Principal Investigator Paper : 03 Module: 14 Principal Investigator Paper Coordinator and Content Writer Dr. Sunil Kumar Khare, Professor, Department of Chemistry, IIT-Delhi Dr. Sunil Kumar Khare, Professor, Department of Chemistry,

More information

Gene Expression Transcription

Gene Expression Transcription Why? ene Expression Transcription How is mrn synthesized and what message does it carry? DN is often referred to as a genetic blueprint. In the same way that blueprints contain the instructions for construction

More information

Unit 6: Gene Activity and Biotechnology

Unit 6: Gene Activity and Biotechnology Chapter 16 Outline The Molecular Basis of Inheritance Level 1 Items students should be able to: 1. Recognize scientists and the experiments that lead to the understanding of the molecular basis of inheritance.

More information

MIT Department of Biology 7.013: Introductory Biology - Spring 2005 Instructors: Professor Hazel Sive, Professor Tyler Jacks, Dr.

MIT Department of Biology 7.013: Introductory Biology - Spring 2005 Instructors: Professor Hazel Sive, Professor Tyler Jacks, Dr. MIT Department of Biology 7.01: Introductory Biology - Spring 2005 Instructors: Professor Hazel Sive, Professor Tyler Jacks, Dr. Claudette Gardel iv) Would Xba I be useful for cloning? Why or why not?

More information

DNA is the genetic material. DNA structure. Chapter 7: DNA Replication, Transcription & Translation; Mutations & Ames test

DNA is the genetic material. DNA structure. Chapter 7: DNA Replication, Transcription & Translation; Mutations & Ames test DNA is the genetic material Chapter 7: DNA Replication, Transcription & Translation; Mutations & Ames test Dr. Amy Rogers Bio 139 General Microbiology Hereditary information is carried by DNA Griffith/Avery

More information

#FD µl (for 200 rxns) Expiry Date: Description. 1 ml of 10X FastDigest Green Buffer. Store at -20 C

#FD µl (for 200 rxns) Expiry Date: Description. 1 ml of 10X FastDigest Green Buffer. Store at -20 C PRODUCT INFORMATION Thermo Scientific FastDigest SalI #FD0644 Lot: 5'...G T C G A C...3' 3'...C A G C T G...5' Supplied with: Store at -20 C 200 µl (for 200 rxns) Expiry Date: BSA included www.thermoscientific.com/onebio

More information

Biotechnology and Energy Conservation. Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman

Biotechnology and Energy Conservation. Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman Biotechnology and Energy Conservation Prof. Dr.oec.troph. Ir. Krishna Purnawan Candra, M.S. Program Magister Ilmu Lingkungan Universitas Mulawarman 12 th Lecture Genetic Engineering The Aim: Students can

More information

Chapter 11 Biotechnology : Principles and Processes. Chapter 12 Biotechnology and Its Applications

Chapter 11 Biotechnology : Principles and Processes. Chapter 12 Biotechnology and Its Applications Chapter 11 Biotechnology : Principles and Processes Chapter 12 Biotechnology and Its Applications Ever since the days of Rene Descartes, the French philosopher, mathematican and biologist of seventeenth

More information