In order to do transformation, the gene to be transferred is placed into a plasmid. This is done with the help of restriction enzymes, 7

Size: px
Start display at page:

Download "In order to do transformation, the gene to be transferred is placed into a plasmid. This is done with the help of restriction enzymes, 7"

Transcription

1 Fluorescent Protein Transformation Student Background Genetic transformation occurs when a cell takes up (i.e. takes inside) and expresses a new piece of genetic material DNA. Genetic transformation literally means change caused by genes and it involves the insertion of a gene(s) into an organism in order to change the organism s traits. Remember that a gene is a piece of DNA which provides the instructions for making (coding for) a particular protein. In this lab, we will transform bacteria, giving it the ability to make fluorescent proteins. These are proteins that absorb light at one wavelength (color) and re-emit the light at a different wavelength, or color. Some things will glow with a new color when you shine the "right light" on them. Most people are accustomed to seeing fluorescence produced by ultraviolet light, often called "black light." Genetic transformation is used in many areas of biotechnology. In agriculture, genes coding for traits such as frost, pest or drought resistance can be genetically transformed into plants. In bioremediation, bacteria (prokaryotes or single-celled organisms without a nucleus) can be genetically transformed with genes enabling them to digest oil spills. A medical application of transformation is in the creation of proteins, such as insulin (synthesized by Genentech) and factor VIII (blood clotting protein synthesized by Bayer). Genes can be cut out of human, animal or plant DNA and placed inside bacteria and the bacteria will produce the foreign protein coded for by the gene. For example, a healthy human gene for the hormone insulin can be put into bacteria. Under the right conditions, these bacteria can make authentic human insulin just as they would make their own proteins. This insulin can then be used to treat patients with the genetic disease, diabetes, whose insulin genes do not function properly. In this lab, you will learn about the process of moving genes form one organism to the other with the aid of a plasmid. In addition to one large circular chromosome which contains all of the genes a bacterium needs for its normal existence, bacteria naturally contain one or more tiny circular pieces of DNA called plasmids. Plasmid DNA contains genes for traits that may be beneficial to bacterial survival under certain environmental conditions. In nature, bacteria can transfer plasmids back and forth, allowing them to share these beneficial genes. This mechanism allows bacteria to adapt to new environments. The recent occurrence of bacterial resistance to antibiotics is due to transmission of plasmids. In order to do transformation, the gene to be transferred is placed into a plasmid. This is done with the help of restriction enzymes, 7

2 naturally occurring enzymes from bacteria that recognize a particular sequence of DNA bases and cut at that sequence. Bacteria use restriction enzymes to protect themselves from viruses which inject their DNA into the bacteria; the enzymes can cut the viral DNA before it can hurt the bacteria. The same restriction enzyme is used to cut the ends of the gene we want to transfer and to cut open the plasmid. Because the cuts are made at the same base sequence, the ends will match and reattach when placed together. To move the plasmid through the E. coli cell membrane you will use a transformation solution of calcium chloride (CaCl 2 ), and a procedure know as heat shock which will allow the plasmid to move through the cell wall and membrane without killing the bacteria. Although no one knows exactly how this works right now, the prevailing hypothesis is that the CaCl 2 causes pores to open in the bacterial cell wall and membrane, allowing the plasmid to enter the cell. The details of this process are one of the things that scientists still have yet to discover. In order to select only the bacteria that have received the new gene, the plasmid also has a gene for resistance to the antibiotic Ampicillin. The gene codes for the production of a protein, beta-lactamase, that allows the bacteria to digest the antibiotic before it can cause any harm. Therefore, if ampicillin is mixed into the agar on the bacterial plates, the only bacteria that can survive there will be bacteria with the gene/plasmid for ampicillin resistance. If any bacteria grow on the plate, they must have been transformed. Each colony on the plate is the offspring of one original transformed bacterial cell (a clone of the original). The bacteria reproduce very rapidly, and a colony may represent millions of cells, all of which are genetically identical, since they all came from the same original bacterium. Below is a map of the plasmid used in this lab. The gene for ampicillin resistance is labeled and FP stands for fluorescent protein. As you can see, any one of the different colored fluorescent proteins can be inserted into the same place in this plasmid. In this lab, each DNA plasmid (1-8) has the gene that codes for a different fluorescent protein. There will be 8 different colored fluorescent plasmid/proteins possible in the class. One of your tasks will be to figure out which protein s instructions were on the plasmid you received. mfruits vector ~ 3.6kb 8

3 Fluorescent Protein Student Background Fluorescent proteins (FPs) are found in light-producing cells of many coelenterates (e.g. jellyfish and corals). In most species their function is an intriguing mystery. But in general in the animal kingdom, bright coloration like this is an adaptation for survival and is used for either finding food by locating or attracting prey, for protection against predators, or for communication between individuals of a given species usually associated with attracting mates. Discosoma (dsred) Fluorescent corals To the left is a picture of the molecular structure of one fluorescent protein (Green Fluorescent Protein or GFP). You can see it has an interesting barrel shape. The part or the molecule that actually fluoresces (the chromophore) is in the middle of the barrel and is composed of a ring of only 3 amino acids. Another interesting thing about fluorescent proteins is that the organism doesn t need to use energy to make the protein light up; the protein is simply energized by light hitting the chromophore. Applications of Fluorescent Proteins FPs can be attached to many proteins without interfering with their functions. If the gene for the FP is attached to a cell s protein gene, then when the normal gene is expressed in the cell, the FP will be expressed at the same time. This makes it possible to visually detect when and where a particular protein of interest is produced and used by a cell. FPs are also being used for screening drugs, evaluating viral vectors for human gene therapy, biological pest control, and monitoring genetically altered microbes in the environment, among other applied efforts. 9

4 One to Two days before the lab: 1. Label eppendorf (microfuge) tubes CaCl 2 and aliquot 1 ml for each lab group. 2. Copy instructions for students 3. Streak starter plates at least one for each class Touch sterile loop to E. coli culture in stab Use sterile loop to streak starter plates, then incubate at 37 o C for 24 hours or 2 days on desktop 4

5 Student Transformation Lab Protocol Period Individuals in my lab group and their assigned tasks: NAME TASK COMPLETED (me) Checklist of materials at each lab station: 1 tube of CaCl 2 on ice 2 empty microtubes 1 waterproof pen 4 disposable transfer pipette (or you can use p20 s with yellow tips) sterile innoculating loops OR sterile tips 2 cotton swabs tape for sealing plates after innoculation 1 LB plate 2 LB/AMP plates cup with ice and water One tube of plasmid labeled either PM1 or PM2 on ice. Class or lab station waste containers with 10% bleach solution. float rack (optional) Checklist for Procedure: 1. Check to be sure you have all the needed materials at your lab station. 2. If your plates have not already been labeled, label one LB/amp plate +, one LB/amp (negative) and LB plate (negative). Be sure to only write near the edge of the plate. 3. Label two microtubes, one with a (+) and one with a ( -) along with your initials and class period. Using a plastic pipette, place 500µl of CaCl 2 (transformation solution) in each tube, close it and place the tube on ice for at least 2 minutes. 4. Using a sterile loop/tip, pick up at least five colonies (5 dots, or part of a streak ) of bacteria from the class starter plate by gently scraping it off from the top of the LB agar on which it is growing. Transfer the colonies to each cold tube of CaCl 2 by swizzling or twisting it around. Mix by inverting the tube to be sure the bacteria are dissolved. The solution should look cloudy with no chunks. If the solution isn t cloudy, you can add more colonies. Place the tubes back on ice. 10

6 5. Using the transfer pipette, add all of the plasmid solution (in small, clear microtube labeled either PM1 or PM2) into the positive (+) tube with CaCl 2 that you had placed on ice. BE SURE THAT THE PLASMID IS ONLY TRANSFERRED TO THE + TUBE. Tap gently to mix. Using a clean transfer pipette, add all of the buffer control to the negative (-) tube. 6. Incubate both tubes on ice for 10 minutes. Make sure the tubes are immersed in the ice water. Tap your tubes gently to mix once or twice during this incubation. 7. Heat shock: Take your ice water bath with its tubes to the 42 o C hot water bath. Transfer the tubes to the hot water and time for exactly 45 seconds. Make sure that the tubes are in contact with the hot water. Immediately return the tube to the ice. 8. Incubate on ice for 2 minutes. 9. Tap your tubes gently to mix. Open the negative (-) tube, and with a clean pipette, transfer 300µL or about half of the cell mixture to the negative (-) LB/amp petri dish you prepared previously. Add the rest of this tube to the LB plate. From the positive (+) tube, and with the same pipette, transfer 300µl or about half of the mixture to ONLY the LB/amp positive (+) plate. 10. Spread the cells on each plate with a clean, sterile loop or cotton swab. Let this sit for a few minutes to absorb the liquid. 11. Stack your plates and tape together. Place in the incubator or follow your teacher s directions. 11

7 Student Pre Lab Questions: Name Period 1. What is the purpose of using ampicillin on one of the plates? 2. What is the purpose of a control in a lab? What was (were) the control(s) in this lab? 3. What genes are present in the plasmid and what is the function of each protein product? 4. In your own words, outline the steps in this procedure and explain the purpose of each step. You may use sketches or cartoons to supplement your answer. (Do this on another piece of paper) 12

8 Results and Discussion Name Period 1. How many fluorescent colonies grew on your plates? What color(s) were they? 2. Use this chart to decide which unknown fluorescent protein your DNA sample codes for. Record your conclusion below the chart. BFP (Y66H) EGFP (S65T) YFP (T203Y) mtangerine mcherry mgrape1 excitation emission 3. Did you have (or see in the classroom) colonies that appeared different in visible v. UV light? Why do you think this might occur? 4. Suppose that a group did not get any transformed colonies. What possible errors could have caused this? 13

9 Student Questions 1. Identify and discuss an ethical issue that was raised in this lab. 2. What is the purpose of developing these fluorescent proteins in biological research? 3. What would you do with fluorescent proteins if it was up to you? 14

pglo Transformation Lab Integrated Science 4 Redwood High School Name Per:

pglo Transformation Lab Integrated Science 4 Redwood High School Name Per: pglo Transformation Lab Integrated Science 4 Redwood High School Name Per: n Introduction To Transformation In this lab you will perform a procedure known as a genetic transformation. Remember that a gene

More information

Bacterial Transformation Using Fluorescent Protein Teacher Guide

Bacterial Transformation Using Fluorescent Protein Teacher Guide Bacterial Transformation Using Fluorescent Protein Teacher Guide sciencebridge PROTOCOL 2 Bacterial Transformation using Fluorescent Protein Central question How does a change in the genotype of an organism

More information

Bacterial Transformation Protocol 2

Bacterial Transformation Protocol 2 26 BACTERIAL TRANSFORMATION USING FLUORESCENT PROTEIN Bacterial Transformation Protocol 2 Group # Role in Group Materials Reader Timer Technician Student Name Materials checklist (1) ScienceBridge Transformation

More information

pglo Transformation 1. Do the genetic transformation. 2. Determine the degree of success in your efforts to genetically alter an organism.

pglo Transformation 1. Do the genetic transformation. 2. Determine the degree of success in your efforts to genetically alter an organism. Introduction to Transformation pg Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides the instructions for making

More information

This lab also contributes to the attainment of the following elements of the 00UK objective:

This lab also contributes to the attainment of the following elements of the 00UK objective: General Biology I The Unity of Life Laboratory Genetic Transformation of Bacteria with pglo 10% of lab mark (2% of final course mark) modified from: BioRad Biotechnology Explorer pglo Bacterial Transformation

More information

Student Manual. pglo Transformation

Student Manual. pglo Transformation Student Manual pglo Transformation STUDENT MANUAL LESSON 1 Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece

More information

Student Manual. pglo Transformation

Student Manual. pglo Transformation Student Manual pglo Transformation STUDENT MANUAL LESSON 1 Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece

More information

Transforming E. Coli with pglo Plasmids

Transforming E. Coli with pglo Plasmids Name: Transforming E. Coli with pglo Plasmids AP Biology Transformation Background: Transformation is a process of transferring genetic information from one organism to another. In bacteria, a small circular

More information

Student Manual. pglo Transformation

Student Manual. pglo Transformation Student Manual pglo Transformation Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides

More information

Student Manual. pglo Transformation

Student Manual. pglo Transformation Student Manual pg Transformation STUDENT MANUA ESSN 1 esson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of

More information

Transformation of E. coli with the Rainbow of BioBridge Fluroescent Plasmids

Transformation of E. coli with the Rainbow of BioBridge Fluroescent Plasmids Transformation of E. coli with the Rainbow of BioBridge Fluroescent Plasmids Introduction: Green fluorescent protein (GFP) was the first fluorescent protein to be discovered and manipulated for use in

More information

ONTARIO SCIENCE CENTRE. Teacher Guide. Way to Glow Program

ONTARIO SCIENCE CENTRE. Teacher Guide. Way to Glow Program ONTARIO SCIENCE CENTRE Teacher Guide Way to Glow Program Table of Contents Bacterial transformation background information 3 Experimental procedure 5 Expected results 7 Post-program activity sheet 8 Post-program

More information

Transforming E. Coli with pglo Plasmids

Transforming E. Coli with pglo Plasmids Name: Transforming E. Coli with pglo Plasmids AP Biology Transformation Background: Transformation is a process of transferring genetic information from one organism to another. In bacteria, a small circular

More information

ONTARIO SCIENCE CENTRE

ONTARIO SCIENCE CENTRE ONTARIO SCIENCE CENTRE Table of Contents Bacterial transformation background information 3 Experimental procedure 5 Expected results 8 Post-program activity sheet 9 Post-program activity sheet with answers

More information

Bacterial Transformation with pglo, Part 1

Bacterial Transformation with pglo, Part 1 Biology 211 Bacterial Transformation with pglo, Part 1 OBJECTIVES: Practice formulating predictions Describe the principles of bacterial transformation. Explain the procedure for gene transfer using plasmid

More information

BACTERIAL TRANSFORMATION LESSON PLAN

BACTERIAL TRANSFORMATION LESSON PLAN BACTERIAL TRANSFORMATION LESSON PLAN Primary Learning Outcomes: Understanding the process of bacterial genetic engineering through plasmid insertion. High School Georgia Performance Standards SCSh2. Students

More information

Biology Lab Activity 4-5 DNA Transformation

Biology Lab Activity 4-5 DNA Transformation Biology Lab Activity 4-5 DNA Transformation Scientists can insert genes into bacteria. The genes inserted in the Indo-Blu process (this lab) are on a circular piece of DNA called a plasmid. (The plasmid

More information

Introduction to pglo lab

Introduction to pglo lab Please take these notes carefully. You do not need to write anything in RED Introduction to pglo lab Bacteria Transformation What is a plasmid? A plasmid is a small circular piece of DNA (about 2,000 to

More information

2 Creating Genetically Modified Bacteria Gl o w - i n-th e - d a r k rabbits, pigs, and mice may sound like something out

2 Creating Genetically Modified Bacteria Gl o w - i n-th e - d a r k rabbits, pigs, and mice may sound like something out 2 Creating Genetically Modified Bacteria Gl o w - i n-th e - d a r k rabbits, pigs, and mice may sound like something out of a science fiction movie, but because of genetic modification, these animals

More information

Bacterial Transformation: Unlocking the Mysteries of Genetic Material

Bacterial Transformation: Unlocking the Mysteries of Genetic Material PR009 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Bacterial Transformation: Unlocking the Mysteries of Genetic Material Teacher s Guidebook

More information

Bacterial Transformation Lab - pglo

Bacterial Transformation Lab - pglo Bacterial Transformation Lab - pglo Name: Date: Pre-Lab Score: Lab Overview: In this investigation, you will gain an understanding of the techniques of culturing E. coli bacteria and transforming using

More information

Exploring STEAM with Transformation

Exploring STEAM with Transformation Exploring STEAM with Transformation Thomas Cynkar Edvotek www.edvotek.com Follow @Edvotek EDVOTEK Biotech The Biotechnology Education Company Celebrating 30 years of science education! EDVOTEK Educatio

More information

You will introduce the GFP gene into the bacterium Escherichia coli using the pglo Bacterial Transformation Kit from Bio-Rad Laboratories.

You will introduce the GFP gene into the bacterium Escherichia coli using the pglo Bacterial Transformation Kit from Bio-Rad Laboratories. Practical 5 - R(a) Bacterial Transformation This practical focuses on- Using complex apparatus, analysis and evaluation Intended learning outcomes By the end of this practical and its write-up you should

More information

pglo mansformation Student Manual Lesson 1 Introduction to Transformation

pglo mansformation Student Manual Lesson 1 Introduction to Transformation Student Manual pglo mansformation Lesson 1 Introduction to Transformation In this lab you will perform a procedure known as genetic transformation. Remember that a gene is a piece of DNA which provides

More information

LABORATORY 5: TRANSFORMING BACTERIA WITH THE LIGATION PRODUCTS

LABORATORY 5: TRANSFORMING BACTERIA WITH THE LIGATION PRODUCTS LABORATORY 5: TRANSFORMING BACTERIA WITH THE LIGATION PRODUCTS So far in your quest to clone a gene you have produced recombinant plasmids and verified that you made the para-r plasmid containing the rfp

More information

Lab 5/5a Transformation of E. coli with a Recombinant Plasmid

Lab 5/5a Transformation of E. coli with a Recombinant Plasmid Lab 5/5a Transformation of E. coli with a Recombinant Plasmid Lab 2 Pre Lab Readiness Familiarity and Proper use of micropipettes Remember the 1 st and 2 nd stops Aseptic Technique Antibiotic Resistance

More information

Lesson 1 Focus Questions Consideration 1: Can I Genetically Transform an Organism? Which Organism?

Lesson 1 Focus Questions Consideration 1: Can I Genetically Transform an Organism? Which Organism? Lesson 1 Focus Questions There are many considerations that need to be thought through in the process of planning a scientific laboratory investigation. Below are a few for you to ponder as you take on

More information

Bacterial Transformation and Protein Purification

Bacterial Transformation and Protein Purification Bacterial Transformation and Protein Purification Group 4 Natalie Beale Gregory A. Pate Justin Rousseau Dohee Won Introduction The purpose of this experiment is to perform a genetic transformation and

More information

Biology 2180 Laboratory #7. Bacterial Growth and Transformation

Biology 2180 Laboratory #7. Bacterial Growth and Transformation Biology 2180 Laboratory #7 Name Bacterial Growth and Transformation Introduction: Most aspects of molecular biology require the use of basic microbiological methods. This section is included for those

More information

Amgen Protocol: Introduction and a few comments:

Amgen Protocol: Introduction and a few comments: Amgen Protocol: Introduction and a few comments: The following is a shortened version of the Amgen Lab. This series of labs involves the creation of a recombinant plasmid, subsequent transformation of

More information

Biotechnology - Transformation Biology Concepts of Biology 7.1

Biotechnology - Transformation Biology Concepts of Biology 7.1 Biotechnology - Transformation Biology 100 - Concepts of Biology 7.1 Name Instructor Lab Section Objectives: To gain a better understanding of: Use of Bacteria in Biotechnology DNA & Plasmid Structure

More information

DNA TRANSFORMATION OF BACTERIA RED COLONY REVISED 3/2003

DNA TRANSFORMATION OF BACTERIA RED COLONY REVISED 3/2003 DNA TRANSFORMATION OF BACTERIA RED COLONY REVISED 3/2003 Prepared by the Office of Biotechnology, Iowa State University TEACHER PREPARATION AND INSTRUCTION GUIDE Preparation for the DNA transformation

More information

Introduction to Genetic Engineering: Bacterial Transformation with Green Fluorescent Protein (pglo)

Introduction to Genetic Engineering: Bacterial Transformation with Green Fluorescent Protein (pglo) Introduction to Genetic Engineering: Bacterial Transformation with Green Fluorescent Protein (pglo) Genetic engineering is an umbrella term that encompasses many different techniques for moving DNA between

More information

Experiment 8: Bacterial Transformation

Experiment 8: Bacterial Transformation Experiment 8: Bacterial Transformation Objectives: At the end of this exercise, you will be able to 1. Understand the concepts of plasmids and genetic transformation. 2. Perform a bacterial transformation

More information

Cloning a Fluorescent Gene

Cloning a Fluorescent Gene Cloning a Fluorescent Gene Laboratory Protocols Handout v1.10 Table of Contents Lab 1: Pipettes and Pipetting... 2 Lab 2: Polymerase Chain Reaction... 5 Lab 3: Ligation... 7 Lab 4: Transformation... 9

More information

Introduction to Genetic Engineering Bacterial Transformation with Green Fluorescent Protein (pglo)

Introduction to Genetic Engineering Bacterial Transformation with Green Fluorescent Protein (pglo) Introduction to Genetic Engineering Bacterial Transformation with Green Fluorescent Protein (pglo) Table of Contents Bacterial Transformation Lab Activity Introduction.....1 Background Information and

More information

Biology 2250 Transformation laboratory

Biology 2250 Transformation laboratory Biology 2250 Transformation laboratory Prior to lab: Discuss how to use micropipettes. Discuss how to use microcentrifuge balance. Discuss how to spread bacteria with alcohol and glass rods. Bacteria are

More information

Feeding a growing world: pglo transformation of E. coli

Feeding a growing world: pglo transformation of E. coli 16 19 2: Transformation Feeding a growing world: pglo transformation of E. coli The issue The Earth s resources are limited, but the human population is growing fast. How can we ensure food security adequate

More information

Figure 1. Map of cloning vector pgem T-Easy (bacterial plasmid DNA)

Figure 1. Map of cloning vector pgem T-Easy (bacterial plasmid DNA) Texas A&M University-Corpus Christi CHEM4402 Biochemistry II Laboratory Laboratory 6: Ligation & Bacterial Transformation (Bring your text and laptop to class if you wish to work on your assignment during

More information

Transformation of E. coli with puc8 Lab Activity Student Study Guide

Transformation of E. coli with puc8 Lab Activity Student Study Guide TM Transformation of E. coli with puc8 Lab Activity Student Study Guide BACKGROUND DID YOU KNOW? Transformation was discovered in the late 1920s by Fred Griffith, an English medical officer, while he was

More information

LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA

LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA LAB #14: Rapid Colony Transformation of E. coli with Plasmid DNA Objective: In this laboratory investigation, plasmids containing fragments of foreign DNA will be used to transform Escherichia coli cells,

More information

Answers to Questions: Teacher s Edition PreLab Activity: Student Learning Outcome & Pre-Lab Predictions For Laboratory Activity

Answers to Questions: Teacher s Edition PreLab Activity: Student Learning Outcome & Pre-Lab Predictions For Laboratory Activity : Teacher s Edition PreLab Activity: Student Learning Outcome & Pre-Lab Predictions For Laboratory Activity Student Learning Outcomes at the end of this laboratory, students will be able to: 1. Explain

More information

Bacterial genetic exchange:transformation

Bacterial genetic exchange:transformation Experiment 11 Laboratory to Biology III Diversity of Microorganisms / Wintersemester / page 1 Experiment Advisor Reading Objectives Background Literature www. Links Bacterial genetic exchange:transformation

More information

Bacterial genetic exchange : Bacterial Transformation

Bacterial genetic exchange : Bacterial Transformation Experiment 11 Laboratory to Biology III: Diversity of Microorganisms 1 Experiment 11 Bacterial genetic exchange : Bacterial Transformation Advisor Munti Yuhana myuhana@botinst.unizh.ch Textbook Chapters

More information

Pacing/Teacher's Notes

Pacing/Teacher's Notes Slide 1 / 31 New Jersey Center for Teaching and Learning Progressive Science Initiative This material is made freely available at www.njctl.org and is intended for the non-commercial use of students and

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

Prepared by the Office of Biotechnology, Iowa State University DRAFT 4/03

Prepared by the Office of Biotechnology, Iowa State University DRAFT 4/03 RECOMBINANT DNA: DUAL ANTIBIOTIC-RESISTANCE GENES Prepared by the Office of Biotechnology, Iowa State University DRAFT 4/03 ** Portions of this protocol were adapted from DNA Science: A First Course in

More information

pglo Worksheet Wednesday, January 31, 2018

pglo Worksheet Wednesday, January 31, 2018 pglo Worksheet Wednesday, January 31, 2018 9:57 PM pglo Worksheet final 2 Page 1 An organism composed of a single cell is better suited for genetic transformation--it can transform at a faster rate - we

More information

AP Biology Lab 6 MOLECULAR BIOLOGY

AP Biology Lab 6 MOLECULAR BIOLOGY AP Biology Laboratory Date: Name and Period: AP Biology Lab 6 MOLECULAR BIOLOGY OVERVIEW In this lab you will investigate some basic principles of molecular biology: 1. Plasmids containing specific fragments

More information

Transduction of an Antibiotic Resistance Gene. Background

Transduction of an Antibiotic Resistance Gene. Background I Student Guide 21-1128 Name------------ Date Transduction of an Antibiotic Resistance Gene Background Transduction is a natural method of gene transfer that occurs in bacteria. The key player in transduction

More information

Yesterday s Picture UNIT 3B

Yesterday s Picture UNIT 3B Warm-Up Plasmids are circular pieces of DNA which bacterial cells are able to take up from the environment, then replicate and transcribe. Eukaryotic cells, by contrast, contain large, linear (non-circular)

More information

Transformation of E. coli with pbr322

Transformation of E. coli with pbr322 The Biotechnology Education Company Revised and Updated Transformation of E. coli with pbr322 EDVO-Kit # 201 Storage: See Page 3 for specific storage instructions Experiment Objective: The objective of

More information

Lab 8: Bacterial Transformation with pglo for Protein Production

Lab 8: Bacterial Transformation with pglo for Protein Production OBJECTIVES: Lab 8: Bacterial Transformation with pglo for Protein Production Describe the principles of chromatography. Explain the procedure for the production of engineered proteins. Isolate the Green

More information

Lesson 1 Introduction to Genetic Engineering

Lesson 1 Introduction to Genetic Engineering Lesson 1 Introduction to Genetic Engineering Genetic engineering is the manipulation of an organism s genetic material (DNA) by introducing or eliminating specific genes A gene is a piece of DNA which

More information

CHAPTER 5A GETTING RECOMBINANT PLASMIDS IN BACTERIA C 35. CHAPTER 5A STUDENT GUIDE 2015 Amgen Foundation. All rights reserved.

CHAPTER 5A GETTING RECOMBINANT PLASMIDS IN BACTERIA C 35. CHAPTER 5A STUDENT GUIDE 2015 Amgen Foundation. All rights reserved. CHAPTER 5A GETTING RECOMBINANT PLASMIDS IN BACTERIA C 35 INTRODUCTION Inserting a gene into a plasmid vector is an important first step in the gene cloning process. However, if the ultimate goal is to

More information

Transformation: Theory. Day 2: Transformation Relevant Book Sections

Transformation: Theory. Day 2: Transformation Relevant Book Sections Day 2: Transformation Relevant Book Sections We will follow the protocols provided in various industry-standard kits, instead of the protocols described in these chapters, but the chapters provide good

More information

How can we use genetic engineering techniques to manipulate heritable information?

How can we use genetic engineering techniques to manipulate heritable information? Genetics and Information Transfer Big Idea 3 INVESTIGATION 8 BIOTECHNOLOGY: BACTERIAL TRANSFORMATION* How can we use genetic engineering techniques to manipulate heritable information? BACKGROUND Are genetically

More information

Mission (Im)possible: Plasmid Mapping Student Materials

Mission (Im)possible: Plasmid Mapping Student Materials Mission (Im)possible: Plasmid Mapping Student Materials Introduction... 2 Pre-Lab Questions... 6 Lab Protocol... 7 Data Collection Worksheet... 11 Post-Lab Questions and Analysis... 12 Last updated: August

More information

Amgen Laboratory Series. Tabs C and E

Amgen Laboratory Series. Tabs C and E Amgen Laboratory Series Tabs C and E Chapter 2A Goals Describe the characteristics of plasmids Explain how plasmids are used in cloning a gene Describe the function of restriction enzymes Explain how to

More information

Molecular Biology for Secondary Classrooms: MBSC

Molecular Biology for Secondary Classrooms: MBSC Molecular Biology for Secondary Classrooms: MBSC Educator and Student Materials unl.mbsc.wordpress.com Program Coordinator: Sarah D. Zulkoski Nebraska EPSCoR epscor.nebraska.edu Contents Module Notes 3

More information

Procedure: GFP cloning project Day 1. Bioinformatics and cloning workshop. Agar plate prep.

Procedure: GFP cloning project Day 1. Bioinformatics and cloning workshop. Agar plate prep. Procedure: GFP cloning project Day 1. Bioinformatics and cloning workshop. Agar plate prep. 1. Prepare nutrient agar (required in next few labs for bacterial work). a. To prepare the agar, weigh 3.85g

More information

Dolan DNA Learning Center Glowing Genes

Dolan DNA Learning Center Glowing Genes Dolan DNA Learning Center Glowing Genes Background History A chromosome is a continuous DNA molecule that can be thousands or millions of base pairs long. The vast length of chromosomes posed a problem

More information

Introduction. Online Resources. 2 VISIT for complete experiment details & free student protocols. Tech Video

Introduction. Online Resources. 2 VISIT   for complete experiment details & free student protocols. Tech Video Transformation Tips and Tricks EDVOTEK WORKSHOP Introduction Are transformations giving you trouble? Then this is the workshop for you! In this workshop, you will transform E. coli with plasmids that express

More information

BIO440 Genetics Laboratory Transformation

BIO440 Genetics Laboratory Transformation BIO440 Genetics Laboratory Transformation The transfer of genetic information between bacteria has been occurring for billions of years. Humans first noticed this process in the laboratory in the 1920

More information

Genetic Engineering: Way to Grow

Genetic Engineering: Way to Grow STO-134 Genetic Engineering: Way to Grow Part 1: Jose s Story Jose is a healthy and active six-year old. The doctor at the health clinic determined that Jose is 35 inches tall. She showed Jose s parents

More information

Purification of mfp. from an Overnight Culture. Laboratory 17

Purification of mfp. from an Overnight Culture. Laboratory 17 Purification of mfp from an Overnight Culture When scientists at a therapeutics company, like Amgen, have successfully identified a promising therapeutic protein, two objectives would be to locate and

More information

Molecular Biology for Secondary Classrooms: MBSC

Molecular Biology for Secondary Classrooms: MBSC Molecular Biology for Secondary Classrooms: MBSC Educator and Student Materials www.unl.mbsc.wordpress.com Program Coordinator: Sarah D. Zulkoski Benson Nebraska EPSCoR www.epscor.unl.edu Contents Module

More information

LAB 1: Eau that smell

LAB 1: Eau that smell LAB 1: Eau that smell Compare 2 competing designs to optimize system performance Acknowledgements: This lab was developed with materials and guidance from the MIT 2006 igem team, as well as technical insights

More information

Recombinant DNA, Biotechnology, and Microbes. Microbiology 221

Recombinant DNA, Biotechnology, and Microbes. Microbiology 221 Recombinant DNA, Biotechnology, and Microbes Microbiology 221 Overview Putting microbes to Work Molecular Cloning Recombinant DNA technology utilizes the power of microbiological selection and screening

More information

MOLEBIO LAB #12: Bacterial Culture Techniques Part I

MOLEBIO LAB #12: Bacterial Culture Techniques Part I MOLEBIO LAB #12: Bacterial Culture Techniques Part I Introduction: This lab introduces an introduction to plating and culturing E. coli on LB agar plates such that single cells can be isolated from one

More information

About Transformation

About Transformation About Transformation In 1928, Frederick Griffith was working on this problem of finding a vaccine against pneumonia caused by the bacteria Streptococcus pneumoniae. Here s what he found: In Experiment

More information

Lab 1 Flow Chart : Learning basic laboratory skills

Lab 1 Flow Chart : Learning basic laboratory skills Lab Flow Chart : Learning basic laboratory skills RD Red dye solution S Dye S2 Dye 2 S3 Dye 3 H 2 O Water X TAE X Lab.: Basic pipetting and serial dilution 2 Plunger button Tip ejector Display window Barrel

More information

Genetic Engineering: Transforming Bacteria

Genetic Engineering: Transforming Bacteria Genetic Engineering: Transforming Bacteria Introduction Activity Introduction In this laboratory experiment, students will transform the phenotype of bacteria through the use of genetic engineering. A

More information

-Is the process of manipulating genes and genomes

-Is the process of manipulating genes and genomes Genetic Engineering -Is the process of manipulating genes and genomes Biotechnology -Is the process of manipulating organisms or their components for the purpose of making useful products Restriction Enzymes

More information

Biotechnology In Your Mouth

Biotechnology In Your Mouth PR005 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Biotechnology In Your Mouth Teacher s Guidebook (Cat. # BE 102) think proteins! think

More information

AP Biology: Unit 5: Development. Forensic DNA Fingerprinting: Using Restriction Enzymes Bio-Rad DNA Fingerprinting Kit

AP Biology: Unit 5: Development. Forensic DNA Fingerprinting: Using Restriction Enzymes Bio-Rad DNA Fingerprinting Kit Forensic DNA Fingerprinting: Using Restriction Enzymes Bio-Rad DNA Fingerprinting Kit Background: Scientists working in forensic labs are often asked to perform DNA profiling or fingerprinting to analyze

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

Isolation & Characterization of Bacteria

Isolation & Characterization of Bacteria PR025 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Isolation & Characterization of Bacteria Teacher s Handbook (Cat. # BE 204) think proteins!

More information

Mission (Im)possible: Determine the Identity of Unknown Plasmids. Student Materials. Introduction Lab Protocol... 5

Mission (Im)possible: Determine the Identity of Unknown Plasmids. Student Materials. Introduction Lab Protocol... 5 Mission (Im)possible: Determine the Identity of Unknown Plasmids Student Materials Introduction... 2 Lab Protocol... 5 Data Collection Worksheet... 9 Pre-Lab Questions... 10 Post-Lab Questions and Analysis...

More information

Bio 121 LAB 11 INSTRUCTIONS - DNA II

Bio 121 LAB 11 INSTRUCTIONS - DNA II Bio 121 LAB 11 INSTRUCTIONS - DNA II In the first part of today's lab we will demonstrate that the DNA which we extracted last week can create heritable changes in the phenotype of bacterial cells. We

More information

GeNei TM Transformation Teaching Kit Manual

GeNei TM Transformation Teaching Kit Manual Teaching Kit Manual Cat No. New Cat No. KT07 107385 KT07A 106220 Revision No.: 00060505 CONTENTS Page No. Objective 3 Principle 3 Kit Description 6 Materials Provided 7 Procedure 9 Observation & Interpretation

More information

Section A: Prokaryotes Types and Structure 1. What is microbiology?

Section A: Prokaryotes Types and Structure 1. What is microbiology? Section A: Prokaryotes Types and Structure 1. What is microbiology? 2. Compare and contrast characteristics of each bacterial type: Eubacteria and Archaebacteria. Eubacteria Both Archaebacteria 3. Label

More information

HiPer Transformation Teaching Kit

HiPer Transformation Teaching Kit HiPer Transformation Teaching Kit Product Code: HTBM017 Number of experiments that can be performed: 10 Duration of Experiment: 4 days Day 1- Preparation of media and revival of E. coli Host Day 2- Inoculation

More information

CRIME SCENE INVESTIGATOR: DNA Profiling

CRIME SCENE INVESTIGATOR: DNA Profiling Bio101- LAB 8 Name: CRIME SCENE INVESTIGATOR: DNA Profiling OBJECTIVES: To review the structure and function of DNA Understand and perform DNA digests To gain experience using the micropipettes and gel

More information

Lab 2C: Basic Techniques. Dilute 10X TE Buffer to Make 1X TE Buffer Determine the Concentration of an Unknown DNA Sample Streak out bacteria colonies

Lab 2C: Basic Techniques. Dilute 10X TE Buffer to Make 1X TE Buffer Determine the Concentration of an Unknown DNA Sample Streak out bacteria colonies Demonstration of sterile technique. Lab 2 Basic Techniques Lab 2A: Lab 2B: Lab 2C: Dilute 10X TE Buffer to Make 1X TE Buffer Determine the Concentration of an Unknown DNA Sample Streak out bacteria colonies

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

Name Per AP: CHAPTER 27: PROKARYOTES (Bacteria) p559,

Name Per AP: CHAPTER 27: PROKARYOTES (Bacteria) p559, AP: CHAPTER 27: PROKARYOTES (Bacteria) p559, 561-564 1. How does the bacterial chromosome compare to a eukaryotic chromosome? 2. What is a plasmid? 3. How fast can bacteria reproduce? 4. What is a bacterial

More information

Cornell Institute for Biology Teachers in partnership with the Pseudomonas-Plant Interaction Project

Cornell Institute for Biology Teachers in partnership with the Pseudomonas-Plant Interaction Project Cornell Institute for Biology Teachers in partnership with the Pseudomonas-Plant Interaction Project Copyright CIBT, 2004. This work may be copied by the original recipient from CIBT to provide copies

More information

Overview: The DNA Toolbox

Overview: The DNA Toolbox Overview: The DNA Toolbox Sequencing of the genomes of more than 7,000 species was under way in 2010 DNA sequencing has depended on advances in technology, starting with making recombinant DNA In recombinant

More information

Unit 2: Metabolism and Survival Sub-Topic (2.7) Genetic Control of Metabolism (2.8) Ethical considerations in the use of microorganisms

Unit 2: Metabolism and Survival Sub-Topic (2.7) Genetic Control of Metabolism (2.8) Ethical considerations in the use of microorganisms Unit 2: Metabolism and Survival Sub-Topic (2.7) Genetic Control of Metabolism (2.8) Ethical considerations in the use of microorganisms Duncanrig Secondary JHM&MHC 2015 Page 1 of 18 On completion of this

More information

1 (a) Define the term genetic engineering [2]

1 (a) Define the term genetic engineering [2] 1 (a) Define the term genetic engineering....[2] (b) Fig. 6.1 is a flow diagram that shows how insulin can be produced using genetic engineering. R Q L M N O P Fig. 6.1 Table 6.1 shows stages in the production

More information

Cell Biology. Sub-Topic (1.5) Genetic Engineering. On completion of this subtopic I will be able to state that

Cell Biology. Sub-Topic (1.5) Genetic Engineering. On completion of this subtopic I will be able to state that Cell Biology Sub-Topic (1.5) Genetic Engineering On completion of this subtopic I will be able to state that Genetic information can be transferred from one cell to another by genetic engineering. Bacteria

More information

Labs 10 and 11: Bacterial Transformation and DNA Purification

Labs 10 and 11: Bacterial Transformation and DNA Purification Biology 107 General Biology Labs 10 and 11: Bacterial Transformation and DNA Purification Molecular biology often involves altering the genetic makeup of an organism in a directed way. In this lab exercise,

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

Who s Your Daddy? Engage: Crime Scene video:

Who s Your Daddy? Engage: Crime Scene video: Who s Your Daddy? 1. Engage: Crime Scene video: Crime Lab Uses DNA to Solve Property Crimes in San Diego County. http://www.youtube.com/watch?v=dxyztbkmxwu Watch the clip and then have groups discuss and

More information

Introduction to Genetic Engineering. Bacterial Transformation with Green Fluorescent Protein (pglo) Teacher Guide

Introduction to Genetic Engineering. Bacterial Transformation with Green Fluorescent Protein (pglo) Teacher Guide Introduction to Genetic Engineering Bacterial Transformation with Green Fluorescent Protein (pglo) Teacher Guide Table of Contents Teacher Guide Letter to Teacher..I Inventory Sheet...II Suggested Transformation

More information

Labs 10 and 11: Bacterial Transformation and DNA Purification

Labs 10 and 11: Bacterial Transformation and DNA Purification Biology 107 General Biology Labs 10 and 11: Bacterial Transformation and DNA Purification Molecular biology often involves altering the genetic makeup of an organism in a directed way. In this lab exercise,

More information

The Biotechnology Education Company. Transformation with Green Fluorescent Protein (GFP) Storage: See Page 3 for specific storage instructions

The Biotechnology Education Company. Transformation with Green Fluorescent Protein (GFP) Storage: See Page 3 for specific storage instructions The Biotechnology Education Company Revised and Updated EDVO-Kit # 223/AP08 Transformation with Green Fluorescent Protein (GFP) Storage: See Page 3 for specific storage instructions EXPERIMENT OBJECTIVE:

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

SYNTHETIC BIOLOGY AND THE HIGH SCHOOL CURRICULUM: LAB 1 _Lab_1

SYNTHETIC BIOLOGY AND THE HIGH SCHOOL CURRICULUM: LAB 1   _Lab_1 SYNTHETIC BIOLOGY AND THE HIGH SCHOOL CURRICULUM: LAB 1 http://openwetware.org/wiki/synthetic_biology_and_the_high_school_curriculum: _Lab_1 LAB 1: Eau that smell Comparing 2 competing designs to optimize

More information