Bacterial Transformation with Green Fluorescent Protein. pgfp Version. Table of Contents Fall 2012

Size: px
Start display at page:

Download "Bacterial Transformation with Green Fluorescent Protein. pgfp Version. Table of Contents Fall 2012"

Transcription

1 Bacterial Transformation with Green Fluorescent Protein pgfp Version Table of Contents Bacterial Transformation Introduction..1 Laboratory Exercise...3 Important Laboratory Practices 3 Protocol Worksheet: Bacterial Transformation Acknowledgements..10

2 Introduction to Bacterial Transformation Transformation is a process of transferring genetic information from one organism to another. In bacteria, a small circular piece of DNA known as a plasmid (Table 1), transfers genetic information between bacteria, allowing these microbes to gain antibiotic resistance and adapt to new environments. This natural process can be modified by humans to increase our quality of life. In agriculture, genes are added to help plants survive difficult climatic conditions, insect damage and increase their nutrients. Toxic chemical spills are often controlled by transformed bacteria. Currently, many diabetics rely on insulin made from bacteria transformed with the human insulin gene. Scientists use transformation as a tool to work on ways to treat other human diseases and conditions. Table 1: Illustration of a bacterial cell with chromosome and plasmids Symbol Bacterial structure Illustration of E. coli Plasmid containing a few genes Circular bacterial chromosome In this lab, you will be using non-pathogenic E. coli bacteria and pgfp, a plasmid modified with two genes. The pgfp plasmid contains the genetic codes for (see Table 2): 1. a green fluorescent protein (GFP) from the bioluminescent jellyfish, Aequorea victoria 2. ampicillin resistance (amp) If pgfp transformation is successful and the bacteria are growing, the colonies will appear green under natural light fluorescent, neon green under UV light. These green bacteria must contain the plasmid with the GFP. For this reason, the green fluorescent protein (GFP) gene is often used as a reporter gene to identify expression of other genes of interest. Table 2: pgfp plasmid and its two important genes. Type of gene Ampicillin resistance gene forming betalactamase, which inactivates ampicillin in media pgfp plasmid with inserted genes GFP% gfp gene produces the green fluorescent protein (GFP) Amp R% 1

3 In order to successfully transform bacteria, you will need to add CaCl 2, transformation solution (TS), to neutralize both the bacterial cell wall and membrane charges, then, quickly shock them with a temperature change in order for them to uptake the pgfp plasmid. After the stressful event, you will provide nutritious broth to restart their growth. Deviating from the protocol listed below may decrease your success in obtaining transformants. Will the untransformed bacteria appear neon green under a UV lamp? Will transformed bacteria fluoresce under a UV lamp? List your predictions below. Before beginning the transformation, observe a plate of E. coli and a vial of pgfp plasmid under a UV lamp. Then, view your transformed colonies once you complete the protocol below. Explain your results. Predictions & Actual Results Item Prediction View with UV Lamp Explanation of Results E. coli growing in petri dish Vial of pgfp plasmid Transformed E. coli in petri dish 2

4 Laboratory Exercise The protocol outlined below describes a procedure for adding plasmid DNA to a bacterial cell. You will cool them quickly and briefly heat the cells to move the plasmid DNA into the cells. Then, you will grow the cells on a petri dish containing LB agar and antibiotics. You will look for the development of green colored colonies of bacteria. Objectives - student should be able to: 1. Understand what a bacterial plasmid is and how it is used in biotechnology. 2. Understand how genes are used to make protein in a cell. a. Add reagents to the bottom of the reaction tube, not to its side. b. Add each additional reagent directly into previously added reagent. c. Do not pipet up and down to mix, as this introduces error. d. Make sure contents are all settled into the bottom of the tube and not on the side or cap of tube. A quick spin may be needed to bring contents down. Important Laboratory Practices a. Pipet slowly to prevent contaminating the pipette barrel. b. Change pipette tips between each delivery. c. Change the tip even if it is the same reagent being delivered between tubes. Change tip every time the pipette is used Keep reagents on ice. Check the box next to each step as you complete it. 3

5 Place a check mark in the box as you complete each step. Protocol 1. Sterilize lab surfaces and wash hands before beginning the lab. 2. Obtain two empty 1.5mL microfuge tubes from your instructor. Using a permanent marker, label one tube +DNA and the other tube DNA. Label each tube twice, on the lid and on the side. Place these tubes into a Styrofoam cup containing crushed ice. 2. Add 250µL of Transformation Solution (TS) to each tube. If using a P-200 micropipettor, set the dial to 125µL and transfer 250µL (125µL, 2 times) Note: TS contains calcium chloride (CaCl 2 ), which helps neutralize both the bacterial cell wall membrane and DNA charges. Keep your tubes on ice. 4. Obtain a starter plate of E. coli. Observe the colonies growing on it and note what you see. 250µL TS DNA Wear safety glasses while using the UV lamp. UV#Light# 5. With a sterile inoculation loop, pick up one bacterial colony from the starter plate. Dip and swirl the loop into the +DNA tube to evenly disperse the colony in the solution and release it from the loop. With the cap closed, flick the tube with your finger to mix. Use a new loop to repeat the process for the - DNA tube. Return tubes to ice. 6. Wearing safety glasses, observe the contents of a vial of pgfp under a UV lamp. 7. With a P-20 micropipettor, transfer 10µL of the pgfp plasmid into your tube labeled +DNA only. DO NOT add plasmid to the DNA tube. UV#Light# Close the cap and flick the tube to mix the plasmid with the contents of this tube. Keep tubes on ice. 250µL pgfp 4

6 8. Incubate both tubes on ice for 10 minutes, making sure the tubes are in contact with the ice. DNA 9. While you re waiting, pick up these 3 plates: 1 LB and 2 LB/amp On the outer edge on bottom of the plate, write +DNA onto one of the LB/amp plates. Write DNA on the LB plate and the second LB/amp plate. Also place your team initial or symbol on the bottom of each plate. 10. Heat shock your bacteria by transferring both tubes to a foam rack and placing them into a water bath set at 42 C for 50 seconds. 10 minutes on ice Plate Name LB DNA LB/amp DNA LB/amp +DNA DNA Make sure the tubes are pushed down as far as they can go in the rack to contact the hot water. DNA DNA After 50 seconds, quickly place both tubes on ice for another 2 minutes. It is VERY important to watch the time and speed of the transfers. 11. Return your tubes to a tube rack now resting on your lab bench. Water&Bath& 42 C%/%50%seconds% 2%min% Add 250µL of LB broth to each of the tubes. If using a P-200 micropipettor, set the dial to 125µL and transfer 250µL (125µL, 2 times) 250µL LB Remember to change the tips between the tubes 12. Close the tubes. Mix each tube by flicking it several times with your finger. Incubate the tubes for at least 10 minutes at room temperature. This process will allow the transformed bacteria to recover and provide nutrients for their growth. 13. Obtain your three labeled plates. Transfer 100µL of the appropriate solution to each labeled plate. Remember to change tips between each transfer 10 minutes at room temperature Plate Name LB DNA LB/amp DNA LB/amp +DNA Add100µL of cell solution 5

7 14. With a new sterile loop for each plate, spread the liquid you just transferred onto each of the plates. Evenly cover as much of the plate as possible. Discard used tips into a waste container with disinfectant. Allow bacteria to saturate into the agar plate for a few minutes before the next step. 15. Invert your three plates. Then stack and tape them together. Place plates into an incubator oven set at 37 C until the next day or when colonies are visible. 16. Decontaminate all lab surfaces with dilute disinfectant and wash hands following the lab 6

8 Name Date Period Worksheet: Bacterial Transformation Lab Predictions Will the untransformed bacteria, pgfp plasmid, and transformed bacteria all fluoresce green? Before viewing these substances with a UV lamp, list your prediction on whether they will fluoresce green. Then, view them under a UV lamp and provide an explanation of your results. 1. Predictions & Results Item Prediction With UV lamp Explanation E. coli colony Vial of pgfp plasmid Transformed E. coli colony 2. Explain the purpose of these processes or substances during transformation. Process or Purpose Substance a. LB agar b. Prevents growth of untransformed bacteria on LB/amp plates. c. Calcium chloride d. Heat shock 3. Describe 2 differences and 2 similarities between these Bacteria. Condition - pgfp DNA bacteria + pgfp DNA bacteria Difference Similarity 7

9 Name Date Period 4. Before transforming your bacteria, list your predictions below for each of these petri dishes and their contents. Then, describe your results following transformation. Contents LB -DNA LB/amp -DNA LB/amp +DNA Predictions* Answers vary Answers vary Answers vary Illustration of Results Description of Results *Possible responses for Predictions - Growth or no growth, fluorescence or no fluorescence under UV light, number of colonies, etc.) 5. Compare your predictions with your actual lab results. Describe how close your predictions were to your actual results and explain possible reasons for any differences. Answers may vary. If responses are different from their results, students may not understand the process and need further review. 6. Explain what may have occurred to produce these results. ( = colony) Contents LB -DNA LB/amp -DNA LB/amp +DNA Illustration of Results Description of Results Possible explanation for results 8

10 Name Date Period 7. If growth appeared on the LB/amp +DNA plate, would these bacteria a. be transformed? Explain. b. fluoresce under UV light? Why or why not? 8. Provide an example of how transformation can be beneficial and an example of how it can be potentially harmful to humans. Condition Transformation example a. Beneficial b. Harmful 9. Provide a rational or benefit of adding DNA sequences coding for fluorescent proteins such as GFP, to tag genes of interest in plasmids used for transformation. 10. In your own words, explain the process of transformation. 9

11 BABEC Educational Transformation Kits For Research Use Only. Not for use in diagnostic procedures. BABEC thanks Qiagen for their generous support of plasmid prep kits for BABEC bacterial transformation labs. 10

Bacterial Transformation with Green Fluorescent Protein. Table of Contents Fall 2012

Bacterial Transformation with Green Fluorescent Protein. Table of Contents Fall 2012 Bacterial Transformation with Green Fluorescent Protein pglo Version Table of Contents Bacterial Transformation Introduction..1 Laboratory Exercise...3 Important Laboratory Practices 3 Protocol...... 4

More information

Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein

Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein INTRODUCTION Green Fluorescent Protein (GFP) is a novel protein produced by the bioluminescent

More information

Transformation of the bacterium E. coli. using a gene for Green Fluorescent Protein

Transformation of the bacterium E. coli. using a gene for Green Fluorescent Protein Transformation of the bacterium E. coli using a gene for Green Fluorescent Protein Background In molecular biology, transformation refers to a form of genetic exchange in which the genetic material carried

More information

GENETIC TRANSFORMATION OF BACTERIA WITH THE GENE FOR GREEN FLUORESCENT PROTEIN (GFP)

GENETIC TRANSFORMATION OF BACTERIA WITH THE GENE FOR GREEN FLUORESCENT PROTEIN (GFP) GENETIC TRANSFORMATION OF BACTERIA WITH THE GENE FOR GREEN FLUORESCENT PROTEIN (GFP) LAB BAC3 Adapted from "Biotechnology Explorer pglo Bacterial Transformation Kit Instruction Manual". (Catalog No. 166-0003-EDU)

More information

Transformation Kit BACTERIAL TRANSFORMATION: GREEN FLUORESCENT PROTEIN. Partnership for Biotechnology and Genomics Education

Transformation Kit BACTERIAL TRANSFORMATION: GREEN FLUORESCENT PROTEIN. Partnership for Biotechnology and Genomics Education Transformation Kit BACTERIAL TRANSFORMATION: GREEN FLUORESCENT PROTEIN Partnership for Biotechnology and Genomics Education Barbara Soots Linda Curro Education Coordinator University of California Davis

More information

LAB 16 Rapid Colony Transformation of E. coli with Plasmid DNA

LAB 16 Rapid Colony Transformation of E. coli with Plasmid DNA LAB 16 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

Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity

Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Today you analyze the results of your bacterial transformation from last week and determine the efficiency

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

Biotechnology Explorer

Biotechnology Explorer arac ori pglo bla GFP Biotechnology Explorer pglo Bacterial Transformation Kit Catalog Number 166-0003EDU explorer.bio-rad.com Store components of this kit at room temperature. Duplication of any part

More information

DNA CAN BE TRANSFERRED BETWEEN BACTERIA GENETIC ENGINEERING USING RECOMBINANT DNA TECHNOLOGY

DNA CAN BE TRANSFERRED BETWEEN BACTERIA GENETIC ENGINEERING USING RECOMBINANT DNA TECHNOLOGY Bacterial Transformation DNA CAN BE TRANSFERRED BETWEEN BACTERIA Background Information Plasmid Transformed Cell Figure 1: Bacterial Transformation Quick Reference Abbreviations GFP pgfp gfp Green fl uorescent

More information

Transferring a Broth Culture to Fresh Broth

Transferring a Broth Culture to Fresh Broth Sterile Technique It is very important in microbiology to work with pure cultures. Unfortunately this is difficult. The world around us is covered with microorganisms. Microorganisms are even carried on

More information

Transformation Protocol

Transformation Protocol To make Glycerol Stocks of Plasmids ** To be done in the hood and use RNase/DNase free tips** 1. In a 10 ml sterile tube add 3 ml autoclaved LB broth and 1.5 ul antibiotic (@ 100 ug/ul) or 3 ul antibiotic

More information

Biotechnology Explorer

Biotechnology Explorer arac ori pglo bla GFP Biotechnology Explorer pglo Bacterial Transformation Kit Catalog #166-0003EDU explorer.bio-rad.com See individual components for storage temperature. Duplication of any part of this

More information

Lab Exercise 3: Media, incubation, and aseptic technique

Lab Exercise 3: Media, incubation, and aseptic technique Lab Exercise 3: Media, incubation, and aseptic technique Objectives 1. Compare the different types of media. 2. Describe the different formats of media, plate, tube etc. 3. Explain how to sterilize it,

More information

GROWING BACTERIA INTRODUCTION

GROWING BACTERIA INTRODUCTION GROWING BACTERIA INTRODUCTION E. coli is a normal part of the bacterial flora of the human gut. It is not generally considered pathogenic, although some strains are highly toxic (recent food poisonings

More information

Transformation of E.coli with pgal

Transformation of E.coli with pgal The Biotechnology Education Company ATTENTION! This experiment includes either BactoBeads or LyphoCells. If you have received LyphoCells, please refer to the addendum posted on the last page of this literature.

More information

Cloning GFP into Mammalian cells

Cloning GFP into Mammalian cells Protocol for Cloning GFP into Mammalian cells Studiepraktik 2013 Molecular Biology and Molecular Medicine Aarhus University Produced by the instructors: Tobias Holm Bønnelykke, Rikke Mouridsen, Steffan

More information

Bacterial Transformation and Plasmid Purification. Chapter 5: Background

Bacterial Transformation and Plasmid Purification. Chapter 5: Background Bacterial Transformation and Plasmid Purification Chapter 5: Background History of Transformation and Plasmids Bacterial methods of DNA transfer Transformation: when bacteria take up DNA from their environment

More information

SAMPLE. Bacterial Transformation. Lab 8 BACKGROUND INFORMATION. Neo/SCI Student s Guide Name... Teacher/Section...

SAMPLE. Bacterial Transformation. Lab 8 BACKGROUND INFORMATION. Neo/SCI Student s Guide Name... Teacher/Section... 1431489 REV 001 Neo/SCI Lab 8 Bacterial Transformation BACKGROUND INFORMATION What Is Biotechnology? Before you start doing biotechnology laboratory exercises, it is important to know exactly what biotechnology

More information

One Shot TOP10 Competent Cells

One Shot TOP10 Competent Cells USER GUIDE One Shot TOP10 Competent Cells Catalog Numbers C4040-10, C4040-03, C4040-06, C4040-50, and C4040-52 Document Part Number 280126 Publication Number MAN0000633 Revision A.0 For Research Use Only.

More information

Biological Sciences Initiative

Biological Sciences Initiative Biological Sciences Initiative HHMI Student Activities Measuring Antibiotic Resistance Introduction: You might be aware that antibiotics were once thought of as a magic bullet; a nearly perfect drug for

More information

LAB 11 PLASMID DNA MINIPREP

LAB 11 PLASMID DNA MINIPREP LAB 11 PLASMID DNA MINIPREP STUDENT GUIDE GOAL The objective of this lab is to perform extraction of plasmid DNA and analyze the results. OBJECTIVES After completion, the student should be able to: 1.

More information

Agrobacterium tumefaciens-mediated transformation of Colletotrichum graminicola and Colletotrichum sublineolum

Agrobacterium tumefaciens-mediated transformation of Colletotrichum graminicola and Colletotrichum sublineolum Agrobacterium tumefaciens-mediated transformation of Colletotrichum graminicola and Colletotrichum sublineolum Flowers and Vaillancourt, 2005. Current Genetics 48: 380-388 NOTE added by L. Vaillancourt:

More information

Quantifying Bacterial Concentration using a Calibrated Growth Curve

Quantifying Bacterial Concentration using a Calibrated Growth Curve BTEC 4200 Lab 2. Quantifying Bacterial Concentration using a Calibrated Growth Curve Background and References Bacterial concentration can be measured by several methods, all of which you have studied

More information

TransformAid Bacterial Transformation Kit

TransformAid Bacterial Transformation Kit Home Contacts Order Catalog Support Search Alphabetical Index Numerical Index Restriction Endonucleases Modifying Enzymes PCR Kits Markers Nucleic Acids Nucleotides & Oligonucleotides Media Transfection

More information

Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual

Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual I. Purpose...1 II. Introduction...1 III. Inhibition of Bacterial Growth Protocol...2 IV. Inhibition of in vitro

More information

The E. coli Insulin Factory

The E. coli Insulin Factory The E. coli Insulin Factory BACKGROUND Bacteria have not only their normal DNA, they also have pieces of circular DNA called plasmids. Plasmids are a wonderfully ally for biologists who desire to get bacteria

More information

UltraClean Soil DNA Isolation Kit

UltraClean Soil DNA Isolation Kit PAGE 1 UltraClean Soil DNA Isolation Kit Catalog # 12800-50 50 preps New improved PCR inhibitor removal solution (IRS) included Instruction Manual (New Alternative Protocol maximizes yields) Introduction

More information

Use of Micropipettes

Use of Micropipettes Use of Micropipettes Prior to lab you should understand: The function of micropipettes in the laboratory Basic parts of micropipette What volumes are measured with P, P and P1 micopipettors How to read

More information

In order to be useful, a smear must have the following qualities:

In order to be useful, a smear must have the following qualities: Smear Preparation and Simple Stain Objectives: Make bacterial smear slides (usually called smears) Distinguish cells on these slides using a simple stain procedure Unstained microbial cells are nearly

More information

Genetics and Information Transfer

Genetics and Information Transfer 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

How Does a Doctor Test for AIDS?

How Does a Doctor Test for AIDS? Edvo-Kit #S-70 How Does a Doctor Test for AIDS? S-70 Experiment Objective: The Human Immunodefi ciency Virus (HIV) is an infectious agent that causes Acquired Immunodefi ciency Syndrome (AIDS) in humans.

More information

UltraClean Forensic DNA Isolation Kit (Single Prep Format)

UltraClean Forensic DNA Isolation Kit (Single Prep Format) UltraClean Forensic DNA Isolation Kit (Single Prep Format) Catalog No. Quantity 14000-10 10 preps 14000-S 1 prep Instruction Manual Please recycle Version: 10302012 1 Table of Contents Introduction...

More information

ExpressArt Bacterial H-TR cdna synthesis kit. With extreme selectivity against rrnas

ExpressArt Bacterial H-TR cdna synthesis kit. With extreme selectivity against rrnas ExpressArt Bacterial H-TR cdna synthesis kit With extreme selectivity against rrnas suitable for 2 to 4 µg total RNA Catalogue No. 8004-A30 (30 rxns) Reagents Materials are provided for 30 cdna synthesis

More information

Catalase. ***You will be working with hot water, acids and bases in this laboratory*** ****Use Extreme Caution!!!****

Catalase. ***You will be working with hot water, acids and bases in this laboratory*** ****Use Extreme Caution!!!**** AP BIOLOGY BIOCHEMISTRY ACTIVITY #9 NAME DATE HOUR CATALASE LAB INTRODUCTION Hydrogen peroxide (H 2 O 2 ) is a poisonous byproduct of metabolism that can damage cells if it is not removed. Catalase is

More information

Module 3: Strawberry DNA Extraction

Module 3: Strawberry DNA Extraction Module 3: Strawberry DNA Extraction Teacher/Leader Target Audience: 7-12 Life Science, Biology, Ag Science Overview: In this lab, students will extract DNA from a strawberry using everyday materials and

More information

RNA Extraction and Quantification, Reverse Transcription, and Real-time PCR (q-pcr)

RNA Extraction and Quantification, Reverse Transcription, and Real-time PCR (q-pcr) RNA Extraction and Quantification, Reverse Transcription, and Real-time Preparation of Samples Cells: o Remove media and wash cells 2X with cold PBS. (2 ml for 6 well plate or 3 ml for 6cm plate) Keep

More information

LAB 4. Cultivation of Bacteria INTRODUCTION

LAB 4. Cultivation of Bacteria INTRODUCTION LAB 4. Cultivation of Bacteria Protocols for use of cultivation of bacteria, use of general growth, enriched, selective and differential media, plate pouring, determination of temperature range for growth

More information

TransIT -2020 Transfection Reagent

TransIT -2020 Transfection Reagent Quick Reference Protocol, MSDS and Certificate of Analysis available at mirusbio.com/5400 INTRODUCTION TransIT -2020 Transfection Reagent is a high-performance, animal-origin free, broad spectrum reagent

More information

LAB 7 DNA RESTRICTION for CLONING

LAB 7 DNA RESTRICTION for CLONING BIOTECHNOLOGY I DNA RESTRICTION FOR CLONING LAB 7 DNA RESTRICTION for CLONING STUDENT GUIDE GOALS The goals of this lab are to provide the biotech student with experience in DNA digestion with restriction

More information

BUGS" THAT PRODUCE DRUGS TO KILL "BUGS Microbes Produce Antibiotics

BUGS THAT PRODUCE DRUGS TO KILL BUGS Microbes Produce Antibiotics BUGS" THAT PRODUCE DRUGS TO KILL "BUGS Microbes Produce Antibiotics Science in the Real World Microbes In Action BUGS" THAT PRODUCE DRUGS TO KILL "BUGS is a curriculum unit developed as part of the Science

More information

Related topics: Application Note 27 Data Analysis of Tube Formation Assays.

Related topics: Application Note 27 Data Analysis of Tube Formation Assays. Tube Formation Assays in µ-slide Angiogenesis Related topics: Application Note 27 Data Analysis of Tube Formation Assays. Contents 1. General Information... 1 2. Material... 2 3. Work Flow Overview...

More information

CLONING IN ESCHERICHIA COLI

CLONING IN ESCHERICHIA COLI CLONING IN ESCHERICHIA COLI Introduction: In this laboratory, you will carry out a simple cloning experiment in E. coli. Specifically, you will first create a recombinant DNA molecule by carrying out a

More information

Disc Diffusion Susceptibility Methods

Disc Diffusion Susceptibility Methods Disc Diffusion Susceptibility Methods Introduction When a filter paper disc impregnated with a chemical is placed on agar the chemical will diffuse from the disc into the agar. This diffusion will place

More information

PREPARATION FOR CHEMISTRY LAB: COMBUSTION

PREPARATION FOR CHEMISTRY LAB: COMBUSTION 1 Name: Lab Instructor: PREPARATION FOR CHEMISTRY LAB: COMBUSTION 1. What is a hydrocarbon? 2. What products form in the complete combustion of a hydrocarbon? 3. Combustion is an exothermic reaction. What

More information

Enzyme Pre-Lab. Using the Enzyme worksheet and Enzyme lab handout answer the Pre-Lab questions the pre-lab must be complete before beginning the lab.

Enzyme Pre-Lab. Using the Enzyme worksheet and Enzyme lab handout answer the Pre-Lab questions the pre-lab must be complete before beginning the lab. Enzyme Pre-Lab Using the Enzyme worksheet and Enzyme lab handout answer the Pre-Lab questions the pre-lab must be complete before beginning the lab. Background: In this investigation, you will study several

More information

Enzymes: Amylase Activity in Starch-degrading Soil Isolates

Enzymes: Amylase Activity in Starch-degrading Soil Isolates Enzymes: Amylase Activity in Starch-degrading Soil Isolates Introduction This week you will continue our theme of industrial microbiologist by characterizing the enzyme activity we selected for (starch

More information

Measuring Cell Viability/Cytotoxicity: Cell Counting Kit-F

Measuring Cell Viability/Cytotoxicity: Cell Counting Kit-F Introduction The Cell Counting Kit-F is a fluorometic assay for the determination of viable cell numbers. Calcein-AM in this kit passes through the cell membrane and is hydrolized by the esterase in the

More information

MEF Starter Nucleofector Kit

MEF Starter Nucleofector Kit page 1 of 7 MEF Starter Nucleofector Kit for Mouse Embryonic Fibroblasts (MEF) MEF display significant phenotypic variations which depend on the strain, the genetic background of the mice they are isolated

More information

HiPer RT-PCR Teaching Kit

HiPer RT-PCR Teaching Kit HiPer RT-PCR Teaching Kit Product Code: HTBM024 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 4 hours Agarose Gel Electrophoresis: 45 minutes Storage Instructions: The

More information

BACTERIAL ENUMERATION

BACTERIAL ENUMERATION BACTERIAL ENUMERATION In the study of microbiology, there are numerous occasions when it is necessary to either estimate or determine the number of bacterial cells in a broth culture or liquid medium.

More information

Plant Genomic DNA Extraction using CTAB

Plant Genomic DNA Extraction using CTAB Plant Genomic DNA Extraction using CTAB Introduction The search for a more efficient means of extracting DNA of both higher quality and yield has lead to the development of a variety of protocols, however

More information

Lab 5: DNA Fingerprinting

Lab 5: DNA Fingerprinting Lab 5: DNA Fingerprinting You are about to perform a procedure known as DNA fingerprinting. The data obtained may allow you to determine if the samples of DNA that you will be provided with are from the

More information

A Guide to Managing Your Biological Waste at the University at Albany

A Guide to Managing Your Biological Waste at the University at Albany A Guide to Managing Your Biological Waste at the University at Albany Section 1 - What you need to know: Definition: "Regulated Medical Waste (RMW) shall mean any of the following waste which is generated

More information

Assessment of Islet Functional Potency by Glucose Stimulated Insulin Secretion

Assessment of Islet Functional Potency by Glucose Stimulated Insulin Secretion Page 1 of 5 PURPOSE: To assess the in vitro functional potency of purified islets by measuring the amount of insulin release upon glucose stimulation. SPECIMEN: Purified human Islet preparation. APPLICABLE

More information

Paper Chromatography: Separation and Identification of Five Metal Cations

Paper Chromatography: Separation and Identification of Five Metal Cations Paper Chromatography: Separation and Identification of Five Metal Cations Objectives Known and unknown solutions of the metal ions Ag +, Fe 3+, Co 2+, Cu 2+ and Hg 2+ will be analyzed using paper chromatography.

More information

NNIN Nanotechnology Education

NNIN Nanotechnology Education NNIN Nanotechnology Education How Quickly Do Bacteria Grow? Teacher s Guide Purpose: Students will relate real-world applications to mathematical concepts by monitoring bacterial growth over one week and

More information

GRS Plasmid Purification Kit Transfection Grade GK73.0002 (2 MaxiPreps)

GRS Plasmid Purification Kit Transfection Grade GK73.0002 (2 MaxiPreps) 1 GRS Plasmid Purification Kit Transfection Grade GK73.0002 (2 MaxiPreps) (FOR RESEARCH ONLY) Sample : Expected Yield : Endotoxin: Format : Operation Time : Elution Volume : 50-400 ml of cultured bacterial

More information

Dot Blot Analysis. Teacher s Guidebook. (Cat. # BE 502) think proteins! think G-Biosciences www.gbiosciences.com

Dot Blot Analysis. Teacher s Guidebook. (Cat. # BE 502) think proteins! think G-Biosciences www.gbiosciences.com PR110 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Dot Blot Analysis Teacher s Guidebook (Cat. # BE 502) think proteins! think G-Biosciences

More information

Welcome to Implementing Inquirybased Microbial Project. Veronica Ardi, PhD

Welcome to Implementing Inquirybased Microbial Project. Veronica Ardi, PhD Welcome to Implementing Inquirybased Microbial Project Veronica Ardi, PhD Microbiology Laboratory Courses CourseSmart: ebook resources http://instructors.coursesmart.com/ Microbiology Laboratory Courses

More information

PRODUCT INFORMATION...

PRODUCT INFORMATION... VIROMER RED In vitro plasmid DNA and mrna Standard Transfection PRODUCT INFORMATION... 2 GENERAL... 2 RED OR YELLOW?... 3 PROTOCOL GUIDELINES... 4 GENERAL REMARKS... 4 CELL CULTURE AND PLATING... 4 FORWARD/REVERSE

More information

Crime Scenes and Genes

Crime Scenes and Genes Glossary Agarose Biotechnology Cell Chromosome DNA (deoxyribonucleic acid) Electrophoresis Gene Micro-pipette Mutation Nucleotide Nucleus PCR (Polymerase chain reaction) Primer STR (short tandem repeats)

More information

Purification of Plasmid DNA

Purification of Plasmid DNA Purification of Plasmid DNA Introduction: The growth of colonies on antibiotic medium provides phenotypic evidence that cells have been transformed. To confirm this at the genotypic level, plasmid DNA

More information

EUROTUBO DELTALAB 4. PETRI DISHES AND LOOPS

EUROTUBO DELTALAB 4. PETRI DISHES AND LOOPS 77 78 90 x 14 mm Petri Dish Made in polystyrene. Vented. Supplied in groups of 20 units, packaged in heat sealed bags. Code 200200 is ASEPTIC. Code 200209 is sterile by gamma radiation. Suitable for automatic

More information

ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols

ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols v.20090807 For research use only. Not for use in diagnostic procedures. Limited License Subject to the terms and conditions of

More information

RESTRICTION ENZYME ANALYSIS OF DNA

RESTRICTION ENZYME ANALYSIS OF DNA University of Massachusetts Medical School Regional Science Resource Center SUPPORTING MATHEMATICS, SCIENCE AND TECHNOLOGY EDUCATION 222 Maple Avenue, Stoddard Building Shrewsbury, MA 01545-2732 508.856.5097

More information

Biotechnology Explorer TM. Green Fluorescent Protein (GFP) Purification Kit. Instruction Manual

Biotechnology Explorer TM. Green Fluorescent Protein (GFP) Purification Kit. Instruction Manual Biotechnology Explorer TM Green Fluorescent Protein (GFP) Purification Kit Instruction Manual Catalog #166-0005EDU explorer.bio-rad.com Duplication of any part of this document is permitted for classroom

More information

BioResearch. RAFT 3D Cell Culture Kit Protocol

BioResearch. RAFT 3D Cell Culture Kit Protocol BioResearch RAFT 3D Cell Culture Kit Protocol RAFT 3D Cell Culture Kit This protocol describes in detail how to produce RAFT 3D Cell Cultures in 96-well plates, 24-well plates and 24-well inserts. It is

More information

AP BIOLOGY 2007 SCORING GUIDELINES

AP BIOLOGY 2007 SCORING GUIDELINES AP BIOLOGY 2007 SCORING GUIDELINES Question 4 A bacterial plasmid is 100 kb in length. The plasmid DNA was digested to completion with two restriction enzymes in three separate treatments: EcoRI, HaeIII,

More information

DETECTION OF BACTERIAL MOTILITY. To demonstrate bacterial motility by microscopic and macroscopic techniques.

DETECTION OF BACTERIAL MOTILITY. To demonstrate bacterial motility by microscopic and macroscopic techniques. DETECTION OF BACTERIAL MOTILITY I. OBJECTIVES To demonstrate bacterial motility by microscopic and macroscopic techniques. To observe flagella in prepared slides stained by specific flagellar stains. II.

More information

DNA Electrophoresis Lesson Plan

DNA Electrophoresis Lesson Plan DNA Electrophoresis Lesson Plan Primary Learning Outcomes: Students will learn how to properly load a well in an agarose gel. Students will learn how to analyze the results of DNA electrophoresis. Students

More information

Aseptic Technique. A GMP/GTP Training Module

Aseptic Technique. A GMP/GTP Training Module Aseptic Technique A GMP/GTP Training Module Aseptic Technique The GMP Facility manufactures products for clinical use These products must meet a number of requirements, one of which is that they are sterile

More information

Policies. Prep Room Policies

Policies. Prep Room Policies Introduction INTRODUCTION The Microbiology Prep Room is located in 531A Life Sciences Building. The telephone number is 372-8609. It is open from 7:30 a.m. to 4:30 p.m. during the fall and spring semesters.

More information

Amazing DNA facts. Hands-on DNA: A Question of Taste Amazing facts and quiz questions

Amazing DNA facts. Hands-on DNA: A Question of Taste Amazing facts and quiz questions 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

More information

Investigating a Eukaryotic Genome: Cloning and Sequencing a Fragment of Yeast DNA

Investigating a Eukaryotic Genome: Cloning and Sequencing a Fragment of Yeast DNA Investigating a Eukaryotic Genome: Cloning and Sequencing a Fragment of Yeast DNA Credits: This lab was created by Sarah C.R. Elgin and developed and written by Kathleen Weston-Hafer. Specific protocols

More information

Science in the Real World Microbes In Action

Science in the Real World Microbes In Action Science in the Real World Microbes In Action Edited by: Teresa Thiel, Ph.D. University of Missouri-St. Louis Program Director & Microbiologist Slick Oil Lab is a curriculum unit developed as part of the

More information

Blood Collection and Processing SOP

Blood Collection and Processing SOP Brisbane Breast Bank Blood Collection and Processing SOP Breast Pathology Laboratory University of Queensland Centre for Clinical Research Blood Collection We collect 30ml of blood from patients who have

More information

Microbiology BIOL 275 DILUTIONS

Microbiology BIOL 275 DILUTIONS DILUTIONS Occasionally a solution is too concentrated to be used as is. For example, when one is performing manual blood counts, the blood contains too many cells to be counted as such. Or when performing

More information

Optimized Protocol sirna Test Kit for Cell Lines and Adherent Primary Cells

Optimized Protocol sirna Test Kit for Cell Lines and Adherent Primary Cells page 1 of 8 sirna Test Kit for Cell Lines and Adherent Primary Cells Kit principle Co-transfection of pmaxgfp, encoding the green fluorescent protein (GFP) from Pontellina p. with an sirna directed against

More information

National Food Safety Standard Food microbiological examination: Aerobic plate count

National Food Safety Standard Food microbiological examination: Aerobic plate count National Food Safety Standard of the People s Republic of China GB4789.2-2010 National Food Safety Standard Food microbiological examination: Aerobic plate count Issued by 2010-03-26 Implemented by 2010-06-01

More information

1. The Determination of Boiling Point

1. The Determination of Boiling Point 1. The Determination of Boiling Point Objective In this experiment, you will first check your thermometer for errors by determining the temperature of two stable equilibrium systems. You will then use

More information

Frozen-EZ Yeast Transformation II Catalog No. T2001

Frozen-EZ Yeast Transformation II Catalog No. T2001 INSTRUCTIONS Frozen-EZ Yeast Transformation II Catalog No. T2001 Highlights High transformation efficiency that yields approximately 10 5-10 6 transformants per μg plasmid DNA (circular). Frozen storage

More information

Instructions. Torpedo sirna. Material. Important Guidelines. Specifications. Quality Control

Instructions. Torpedo sirna. Material. Important Guidelines. Specifications. Quality Control is a is a state of the art transfection reagent, specifically designed for the transfer of sirna and mirna into a variety of eukaryotic cell types. is a state of the art transfection reagent, specifically

More information

A and B are not absolutely linked. They could be far enough apart on the chromosome that they assort independently.

A and B are not absolutely linked. They could be far enough apart on the chromosome that they assort independently. Name Section 7.014 Problem Set 5 Please print out this problem set and record your answers on the printed copy. Answers to this problem set are to be turned in to the box outside 68-120 by 5:00pm on Friday

More information

Creatine Kinase (CK) Enzymatic Assay Kit Manual Catalog #: 3460-07

Creatine Kinase (CK) Enzymatic Assay Kit Manual Catalog #: 3460-07 Creatine Kinase (CK) Enzymatic Assay Kit Manual Catalog #: 3460-07 TABLE OF CONTENTS GENERAL INFORMATION... 2 Product Description... 2 Procedure Overview... 2 Kit Contents, Storage and Shelf Life... 3

More information

NIH Mammalian Gene Collection (MGC)

NIH Mammalian Gene Collection (MGC) USER GUIDE NIH Mammalian Gene Collection (MGC) Catalog number FL1002 Revision date 28 November 2011 Publication Part number 25-0610 MAN0000351 For Research Use Only. Not for diagnostic procedures. ii Table

More information

MGC premier Expression-Ready cdna clones TCH1103, TCM1104, TCR1105, TCB1106, TCH1203, TCM1204, TCR1205, TCB1206, TCH1303, TCM1304, TCR1305

MGC premier Expression-Ready cdna clones TCH1103, TCM1104, TCR1105, TCB1106, TCH1203, TCM1204, TCR1205, TCB1206, TCH1303, TCM1304, TCR1305 MGC premier Expression-Ready cdna clones TCH1103, TCM1104, TCR1105, TCB1106, TCH1203, TCM1204, TCR1205, TCB1206, TCH1303, TCM1304, TCR1305 The MGC premier Expression-Ready collection has the high quality,

More information

Fighting the Battles: Conducting a Clinical Assay

Fighting the Battles: Conducting a Clinical Assay Fighting the Battles: Conducting a Clinical Assay 6 Vocabulary: In Vitro: studies in biology that are conducted using components of an organism that have been isolated from their usual biological surroundings

More information

LAB 14 ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA)

LAB 14 ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) STUDENT GUIDE LAB 14 ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) GOAL The goal of this laboratory lesson is to explain the concepts and technique of enzyme linked immunosorbent assay (ELISA). OBJECTIVES

More information

Procedure for RNA isolation from human muscle or fat

Procedure for RNA isolation from human muscle or fat Procedure for RNA isolation from human muscle or fat Reagents, all Rnase free: 20% SDS DEPC-H2O Rnase ZAP 75% EtOH Trizol Chloroform Isopropanol 0.8M NaCitrate/1.2M NaCl TE buffer, ph 7.0 1. Homogenizer-probe

More information

UTILIZATION of PLASMA ACTIVATED WATER in Biotechnology, Pharmacology and Medicine. JSC TECHNOSYSTEM-ECO Moscow, Russia April, 2009

UTILIZATION of PLASMA ACTIVATED WATER in Biotechnology, Pharmacology and Medicine. JSC TECHNOSYSTEM-ECO Moscow, Russia April, 2009 UTILIZATION of PLASMA ACTIVATED WATER in Biotechnology, Pharmacology and Medicine JSC TECHNOSYSTEM-ECO Moscow, Russia April, 2009 METHOD of WATER ACTIVATION with PLASMA of GAS DISCHARGE ANODE VACUUM WATER

More information

Transfection reagent for visualizing lipofection with DNA. For ordering information, MSDS, publications and application notes see www.biontex.

Transfection reagent for visualizing lipofection with DNA. For ordering information, MSDS, publications and application notes see www.biontex. METAFECTENE FluoR Transfection reagent for visualizing lipofection with DNA For ordering information, MSDS, publications and application notes see www.biontex.com Description Cat. No. Size METAFECTENE

More information

Lab Safety and Standard Operating Procedures. Faculty of Dentistry And School of Biomedical Engineering

Lab Safety and Standard Operating Procedures. Faculty of Dentistry And School of Biomedical Engineering Lab Safety and Standard Operating Procedures Faculty of Dentistry And School of Biomedical Engineering Introduction It is the requirement that students working in research laboratories at Dalhousie have

More information

DNA SPOOLING 1 ISOLATION OF DNA FROM ONION

DNA SPOOLING 1 ISOLATION OF DNA FROM ONION DNA SPOOLING 1 ISOLATION OF DNA FROM ONION INTRODUCTION This laboratory protocol will demonstrate several basic steps required for isolation of chromosomal DNA from cells. To extract the chromosomal DNA,

More information

Your Best Choice For Laboratory Consumables

Your Best Choice For Laboratory Consumables Your Best Choice For Laboratory Consumables ISO9001:2008 Certificate No. 34730 Report No.01893 Registration No. 10035061 Wuxi NEST Biotechnology., LTD Headquarters Add: No 230, Xida Road, New District,

More information

Biology 3A Laboratory: Enzyme Function

Biology 3A Laboratory: Enzyme Function Biology 3A Laboratory: Enzyme Function Objectives To be able to list the general characteristics of enzymes. To study the effects of enzymes on the rate of chemical reactions. To demonstrate the effect

More information

PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200

PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200 3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System)

More information

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY GENE CLONING AND RECOMBINANT DNA TECHNOLOGY What is recombinant DNA? DNA from 2 different sources (often from 2 different species) are combined together in vitro. Recombinant DNA forms the basis of cloning.

More information

Microbiological Testing of the Sawyer Mini Filter. 16 December 2013. Summary

Microbiological Testing of the Sawyer Mini Filter. 16 December 2013. Summary Microbiological Testing of the Sawyer Mini Filter 16 December 2013 Summary The Sawyer Mini Filter was tested for its ability to remove three microorganisms Raoultella terrigena, Bacillus subtilis, and

More information

The fastest spin-column based procedure for purifying up to 10 mg of ultra-pure endotoxin-free transfection-grade plasmid DNA.

The fastest spin-column based procedure for purifying up to 10 mg of ultra-pure endotoxin-free transfection-grade plasmid DNA. INSTRUCTION MANUAL ZymoPURE Plasmid Gigaprep Kit Catalog Nos. D4204 (Patent Pending) Highlights The fastest spin-column based procedure for purifying up to 10 mg of ultra-pure endotoxin-free transfection-grade

More information