ASEPTIC TRANSFER & PURE CULTURE TECHNIQUES

Save this PDF as:

Size: px
Start display at page:

Download "ASEPTIC TRANSFER & PURE CULTURE TECHNIQUES"

Transcription

1 ASEPTIC TRANSFER & PURE CULTURE TECHNIQUES GENERAL GUIDELINES & REMINDERS: SAFETY: NO EATING OR DRINKING IN THE LAB! Wash your hands with soap both BEFORE and AFTER lab, and, in addition, when you have a spill. Be sure to have nonessential materials (the ONLY essential thing is the lab handouts and a notebook to write in) off of the table. ALWAYS disinfect the tables BEFORE and AFTER lab. ALWAYS check agar plates carefully to make sure that there are no mold or bacterial contaminants on the plate: if so, discard the plate in the autoclave bag. Do the same with any tubed media that you pick up: if contaminated, discard the tube in the discard racks. If there is a spill: o Tell your instructor about it. o Flood the area of the spill with disinfectant and leave on for 10 minutes before using paper towels to soak up. The gas should be turned all of the way on, so that the level is parallel with the rubber tubing. THE LAST PERSON ON THE TABLE CHECKS TO MAKE SURE THAT ALL 4 GAS JETS ARE OFF! INOCULATION, INCUBATION AND LABELING OF CULTURES: You always pick up your microorganisms as a set for your table, sharing them between the table members, and replace them in the biosafety hood. Keep test tube caps and petri dish covers on media to reduce contamination (matters not whether it is sterile media or already cultured). If you see water running on the agar plate, you can do 2 things: o Place the agar plate upside down in the 37C incubator with the top cracked until dry.. o Get another agar plate. Place test tubes in racks when working at your table: never lay the tubes down they leak. Heat the entire piece of metal of the inoculation instrument in the flame: it should be RED HOT. Be sure to COOL your inoculation instrument before picking the inoculum. Use an inoculating needle for agar deeps and an inoculating loop for agar plates and broths. ALWAYS use the proper aseptic technique when transferring cultures from one medium to another. Be sure to put any extra media tubes BACK into the racks if unused: Replace agar plates back into bags if not used. Label all test tubes and petri plates with your name (initials), your table #, date, exercise #, and name of organism. Use masking tape to mark on for the tubes: You can use tape or mark directly on the plates. DISPOSAL OF CULTURES: Do not dump ANY microbial suspension down the drain only on the discard cart. REMOVE ALL labels from test tubes when putting them on the discard cart. Tubes for disposal go inn test tube racks at disposal area in corner of the lab. Agar plates for disposal go into the red biohazard autoclave bag on the floor below the tubes.

2 All agar plates are incubated UPSIDE DOWN (exceptions will be pointed out occasionally). WHY? 1. It reduces bacterial contamination since the bacteria, even if they get into the plate in between the lid and bottom, would have to go UP to get to the agar. 2. It reduces the possibility of water condensation that may be on the lid dropping onto the agar, causing fluid to run across the agar medium. By now you know what media is: it is the nutrient-rich material that provides food to the microbes. There are 3 forms of media: An agar medium contains agar (1.5-2%) as a solidifying agent---isolation and purification. A broth medium has no agar: it is for fast, luxuriant growth. A semi-solid has a reduced percentage of agar, and, therefore, has qualities of both types. In addition to nutrients and growth factors, and perhaps agar, there are additives such as NaCl salt, ph buffers, and ph indicators that allow biochemical reactions to be identified. In this exercise you will learn how to subculture bacteria, using a variety of culture media as your inocula sources and as your new culture media. Different species of bacteria will be used so that you can become familiar with different growth patterns. You will also have a mixed culture with 2 species in order to learn how to best separate and isolate bacterial species. THE OBJECTIVES: Subculture bacteria in/on sterile media of various forms. Eliminate potential contamination of bacterial cultures by using aseptic technique. Practice hand coordination required in good transfer techniques. Identify different ways by which bacteria grow in culture in agar deeps, on agar slants, on agar plates, in broths. Streak out cultures for isolation and identify colonies. MATERIALS NEEDED: set of cultures for the table: a TSA (trypticase soy agar) slant culture of Bacillus subtilis a TSB (trypticase soy broth) culture of Staph epidermidis a TSA plate culture of E. coli a culture of E. coli and Staphylococcus mixed together sterile media: (per person) 3 TSB 2 TSA slants 2 TSA plates THE PROCEDURES: per person There are descriptions how to inoculate an agar plate, a broth tube, an agar slant, and so on below.

3 THE TRANSFERS YOU WILL BE DOING IN LAB: Use an inoculating loop for the agar plate and the broth media. Subculture a broth culture of Staph epidermidis onto a TSA slant Subculture a slant culture of Bacillus subtilus into TSB. Subculture TSA plate culture of E. coli into TSB, a TSA slant, and a TSA plate. Subculture a mix of E. coli and Staphylococcus in TSB and on a TSA plate Although your instructor will show you how to perform these procedures, here are the methods you will use. ASEPTIC TECHNIQUE: 1. Have both the culture that you are taking the inoculum from and the new, sterile medium in front of you. Be sure that the new medium is already labeled so you do not confuse the various cultures. 2. Pick up both tubes in the hand not using the inoculation instrument. 3. Heat the inoculating wire of the loop until red- hot, and be sure that the ENTIRE wire is sterilized. You are now ready to pick the inoculum from the bacterial culture. 4. Keeping the sterile inoculation instrument in your hand, remove both tube caps with your little finger. 5. Run the tops of the tubes through the heat to create an updraft (taking air contaminants AWAY from the tube entrance). 6. Go into the tube to take your inoculum and QUICKLY place the inoculum into the new medium tube. 7. Sterilize the tops of the tubes again (to eliminate potential air contamination again) and replace the caps. 8. Incubate the plates and tubes in the 30 degree C incubator. Look at the section below in INTERPRETION to read your tube results.

4 TAKING THE INOCULUM: FROM A BROTH CULTURE: The inoculum is obtained by first shaking the culture a bit, and then going into the broth with the loop. A film of broth culture can be seen across the loop as you remove it from the tube. FROM AN AGAR SLANT CULTURE: The inoculum is picked off of the top of the slant, like a scraping motion. FROM AN AGAR PLATE CULTURE: The plate cultures have isolated colonies of bacteria growing on them. Pick only one colony or a bit of a colony, if big, with your loop. Lift the lid of the plate just a bit, in order to get your inoculating loop into it: DO NOT TAKE THE ENTIRE LID OFF. INOCULATING THE NEW MEDIUM TO A BROTH CULTURE: The inoculum is just knocked around in the broth, and against the sides of the tube in the broth. TO AN AGAR SLANT CULTURE: Place the loop with the bacteria into the slant tube, all the way down to the bottom of the slant. There are 2 ways to inoculate the slant: If your goal is to identify the type of growth pattern, then just bring the loop straight up the slant. If your goal is to have a luxuriant culture, inoculate in a zig-zag pattern, starting at the bottom of the slant. This increases the surface area of the culture. TO AN AGAR PLATE CULTURE: see directions below on streak technique TO AN AGAR DEEP: Use the needle to inoculate the deeps or semi-solid agars. Stab the inoculum down to the bottom of the deep in a clean, straight stroke. STREAKING AN AGAR PLATE: Until you become well-acquainted with this procedure, you might want to draw the 3 sections that you will streak inside of, on the back (bottom of plate containing agar medium) with a sharpie pen. Keep in mind that you flame your inoculating loop between each section streaked.

5 1. Pick up a loopful of your inoculum from either a broth or an agar culture. Using a sterile agar medium plate (lift the lid just enough to insert the loop), streak a vertical line straight down. 2. When streaking the agar, keep the loop horizontal and only streak the surface of the agar: DO NOT DIG INTO THE AGAR. 3. Move the loop in a zig-zag pattern across the agar until 1/3 of the plate is covered, finishing the first section. 4. Sterilize the loop in the flame and let it cool before continuing to spread the bacteria. You can do this by 1) sticking the hot loop in the agar at the edge of the agar away from the bacteria, or 2) just holding the loop for a few seconds while it cools. 5. Rotate the plate about 90 degrees and spread the bacteria from the first streak into a second area using the same zig-zag spread technique. 6. Sterilize the loop again. Rotate the plate about 90 degrees and spread the bacteria from the second streak into the 3rd area in the same pattern. 7. Sterilize the loop again. Replace the lid and invert the plate. Incubate the plate. INTERPRETATION OF RESULTS: AFTER INCUBATION, check the growth patterns of all tubes and plates.

6 GROWTH PATTERN ON AN ISOLATION STREAK PLATE You should see bacterial cells growing along streak lines and in isolated areas.

7 QUESTIONS: 1. Why streak from the bottom of the agar slant medium up to the top in a straight line, rather than a back and forth wavy inoculation from side-to-side on the slant? 2. Of what use is it to know what kind of growth pattern on an agar slant or a broth medium an organism has? 3. Why do you cross over back the 2nd streak section back into the 1st section, and from the 3rd section back across the 2nd section? 4. Why use a streak plate to grow a bacterium rather than an agar medium slant or a broth medium? Fall Jackie Reynolds, Richland College, Biol 2421

TRANSFER OF BACTERIA USING ASEPTIC TECHNIQUE

TRANSFER OF BACTERIA USING ASEPTIC TECHNIQUE TRANSFER OF BACTERIA USING ASEPTIC TECHNIQUE GENERAL GUIDELINES: Safety Wear a lab coat and have your goggles on! ALWAYS disinfect the tables BEFORE and AFTER lab. Wash your hands with soap both BEFORE

More information

PURE CULTURE TECHNIQUES

PURE CULTURE TECHNIQUES PURE CULTURE TECHNIQUES Most specimens (from animal tissue, plant tissue, or environmental samples) will be mixed, with a variety of bacteria (or other microorganisms). A single gram of feces, for example,

More information

Aseptic Techniques. A. Objectives. B. Before coming to lab

Aseptic Techniques. A. Objectives. B. Before coming to lab Aseptic Techniques A. Objectives Become familiar with 1. The ubiquity of microorganisms (see Note 1) 2. Aseptic techniques (see Note 2) 3. Standard methods for growing/observing microorganisms (see Note

More information

LABORATORY #2 -- BIOL 111 BACTERIAL CULTIVATION & NORMAL FLORA

LABORATORY #2 -- BIOL 111 BACTERIAL CULTIVATION & NORMAL FLORA LABORATORY #2 -- BIOL 111 BACTERIAL CULTIVATION & NORMAL FLORA OBJECTIVES After completing this exercise you should be able to: 1. Identify various types of media 2. Isolate bacteria using aseptic technique.

More information

ENVR 421 Laboratory #1: Basic Bacteriology Techniques

ENVR 421 Laboratory #1: Basic Bacteriology Techniques ENVR 421 Laboratory #1: Basic Bacteriology Techniques Introduction The purpose of this laboratory exercise is to familiarize you with two fundamental bacteriology techniques: the streak plate and the spread

More information

LAB NOTES FOR EXAM 1 SECTION

LAB NOTES FOR EXAM 1 SECTION LAB NOTES FOR EXAM 1 SECTION EX. 2-1: DIVERSITY AND UBIQUITY OF MICROOGANISMS Purpose: Microorganisms are found everywhere in the environment around us. To demonstrate this and to get a taste of the different

More information

Microbiological Methods

Microbiological Methods Microbiological Methods Making Media Pouring Culture Plates Sterile Technique Inoculating Plates and Culture Tubes Use of a Plate Counter to Estimate Microbial Population Densities Sterile Technique Sterile

More information

Microbiological Methods

Microbiological Methods Microbiological Methods Making Media Pouring Culture Plates Sterile Technique Inoculating Plates and Culture Tubes Use of a Plate Counter to Estimate Microbial Population Densities Culturing Microorganisms

More information

Bacterial Plate Preparation. ~ Using aseptic techniques ~

Bacterial Plate Preparation. ~ Using aseptic techniques ~ Bacterial Plate Preparation ~ Using aseptic techniques ~ Bacterial Plates Laboratory and research scientists have to prepare nutrient media to grow specific strains of bacteria for their research. To do

More information

Exercise 4 ASEPTIC TECHNIQUE & STREAK PLATE PREPARATION

Exercise 4 ASEPTIC TECHNIQUE & STREAK PLATE PREPARATION Introduction Exercise 4 ASEPTIC TECHNIQUE & STREAK PLATE PREPARATION In order to work with pure microbial cultures, microbiologists must start with sterile culture media and must be able to prevent contamination.

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

STUDENT PAGE. Using Sterile Technique to Inoculate Bacterial Plates

STUDENT PAGE. Using Sterile Technique to Inoculate Bacterial Plates Using Sterile Technique to Inoculate Bacterial Plates A sterile, or aseptic, technique is used to prevent microbial organisms from contaminating any surface other than the specific location where they

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

COUNT METHOD 5.0 OBJECTIVES 5.1 INTRODUCTION 5.2 PRINCIPLE. Structure

COUNT METHOD 5.0 OBJECTIVES 5.1 INTRODUCTION 5.2 PRINCIPLE. Structure Food Microbiology EXPERIMENT 5 STANDARD PLATE COUNT METHOD Structure 5.0 Objectives 5.1 Introduction 5.2 Principle 5.3 Materials Required 5.4 Procedure 5.4.1 E-coli Culture 5.4.2 Food Samples 5.5 Observations

More information

Microorganisms In Our Environment

Microorganisms In Our Environment PR015 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Microorganisms In Our Environment Teacher s Guidebook (Cat. # BE 106) think proteins!

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

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

SCHEDULE. Friday: Pet Investigations: Plate counts - how to know how many clones of your pet you have (pg. 9-10)

SCHEDULE. Friday: Pet Investigations: Plate counts - how to know how many clones of your pet you have (pg. 9-10) SCHEDULE Wednesday: Pet Investigations: Phenol Red Broth with Durham tubes (pg. 3-4) Oxidation/Fermentation Agar (pg. 5-6) Anaerobic Growth (pg. 7) Growth in Liquid Culture (pg. 8-9) Friday: Pet Investigations:

More information

GENUS STAPHYLOCOCCUS: Isolation and Identification

GENUS STAPHYLOCOCCUS: Isolation and Identification GENUS STAPHYLOCOCCUS: Isolation and Identification Staphylococcus is a genus of Gram +, nonspore-forming cocci belonging to the family Micrococcaceae that are often found as normal human microbiota of

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

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

WHY DO THEY PUT MINT IN TOOTHPASTE? WOULD GARLIC BE BETTER?

WHY DO THEY PUT MINT IN TOOTHPASTE? WOULD GARLIC BE BETTER? Activity 4.22 Student Sheet WHY DO THEY PUT MINT IN TOOTHPASTE? WOULD GARLIC BE BETTER? Purpose To investigate the antibacterial properties of plants. To develop practical skills. YOU NEED Agar plate seeded

More information

Lab Exercise #4 Microbial Control Lab Exercise #4 Control of Microorganisms: Physical, Chemical and Chemotherapeutic

Lab Exercise #4 Microbial Control Lab Exercise #4 Control of Microorganisms: Physical, Chemical and Chemotherapeutic Lab Exercise #4 Control of Microorganisms: Physical, Chemical and Chemotherapeutic I. OBJECTIVES: Investigate the effectiveness various agents of control. Assess the effectiveness of heat in killing vegetative

More information

SELECTED QUESTIONS F ROM OLD MICRO 102 QUIZZES PART I EXPERIMENTS 1 THROUGH 7

SELECTED QUESTIONS F ROM OLD MICRO 102 QUIZZES PART I EXPERIMENTS 1 THROUGH 7 SELECTED QUESTIONS F ROM OLD MICRO 102 QUIZZES PART I EXPERIMENTS 1 THROUGH 7 Question numbers refer to the applicable experiment. Questions with blanks are multiple true-false questions unless otherwise

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

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

CULTURING MICROORGANISMS

CULTURING MICROORGANISMS CULTURING MICROORGANISMS Question practice Name: Class: Date: Time: 24 minutes Marks: 24 marks Comments: BIOLOGY ONLY Page of 2 The diagram shows a method used to grow pure cultures of a bacterium. (a)

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 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

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

How to perform a Gram Stain. Jasleen Singh

How to perform a Gram Stain. Jasleen Singh How to perform a Gram Stain Jasleen Singh Table of Contents iii Table of Contents Table of Contents... iii Introduction... 5 Terminology... 7 Terms to be familiar with... 7 Gram Staining... 8 What is

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

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

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

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

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

M. Dalbey/Bio 105M Isolation of E. coli - Isolation of E. coli from an Environmental Sample

M. Dalbey/Bio 105M Isolation of E. coli - Isolation of E. coli from an Environmental Sample Isolation of E. coli from an Environmental Sample We want to expand our horizons a bit beyond the domesticated lab strains of E. coli. In this exercise you will isolate "wild" E. coli strains from an environmental

More information

LAB NOTES FOR EXAM 1 SECTION

LAB NOTES FOR EXAM 1 SECTION LAB NOTES FOR EXAM 1 SECTION EX. 2-1: DIVERSITY AND UBIQUITY OF MICROOGANISMS Purpose: Microorganisms are found everywhere in the environment around us. To demonstrate this and to get a taste of the different

More information

Making Saline SOLUTION. Lab Number 2 Part 1

Making Saline SOLUTION. Lab Number 2 Part 1 Making Saline SOLUTION Lab Number 2 Part 1 Purpose The purpose of part 1 of this lab is to learn the proper way to make reagents that are needed for labs. Materials Need for the Lab are: Volumetric flasks

More information

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 Division of Environmental Health Sciences Albany, New York NYS DOH LEB-608 Identification

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

number Done by Corrected by Doctor

number Done by Corrected by Doctor L number Lab 2 Done by حسام أبو عوض Corrected by Mahdi sharawi Doctor In many cases we need to identify the type of bacteria causing an infection in order to be able to choose the right medication (antibiotic).

More information

The ramylase Project by Ellyn Daugherty

The ramylase Project by Ellyn Daugherty PR078 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name The ramylase Project by Ellyn Daugherty Transformation of E. coli with pamylase (Lab

More information

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 Division of Environmental Health Sciences Albany, New York NYS DOH LEB-612 Identification

More information

Exercise 13 DETERMINATION OF MICROBIAL NUMBERS

Exercise 13 DETERMINATION OF MICROBIAL NUMBERS Exercise 13 DETERMINATION OF MICROBIAL NUMBERS Introduction When biologists discuss the growth of microorganisms (microbial growth), they are actually referring to population size rather than to the size

More information

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 Division of Environmental Health Sciences Albany, New York NYS DOH LEB-610 Identification

More information

BIOSAFETY GUIDELINES BACKGROUND ON BIOSAFETY

BIOSAFETY GUIDELINES BACKGROUND ON BIOSAFETY BIOSAFETY GUIDELINES BACKGROUND ON BIOSAFETY In addition to following general laboratory safety rules, additional rules must be implemented in the microbiology lab since students are working with living

More information

Lab Exercise 13: Growth Curve

Lab Exercise 13: Growth Curve Lab Exercise 13: Growth Curve OBJECTIVES 1. Know the different phases of a standard growth curve. 2. Understand and perform direct measurement of bacterial growth through serial dilutions and standard

More information

Culturing microorganisms may be hazardous

Culturing microorganisms may be hazardous Practical 8 - S(d) The Effect of Penicillin on Bacterial Growth In this practical focuses on the practical skills of: Planning defining the problem You will be developing other assessed skills throughout

More information

M I C R O B I O L O G Y

M I C R O B I O L O G Y ninth edition TORTORA FUNKE CASE M I C R O B I O L O G Y a n i n t r o d u c t i o n 6 Microbial Growth PowerPoint Lecture Slide Presentation prepared by Christine L. Case Microbial Growth Microbial growth

More information

Project 7: Wound Cultures and Identification

Project 7: Wound Cultures and Identification Project 7: Wound Cultures and Identification Readings: https://labtestsonline.org/understanding/analytes/wound-culture/tab/test Identification of Gram-Positive & Gram-Negative Bacteria Guide to laboratory

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

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

EXPERIMENT. Environmental Influences on Microbial Growth Salt Tolerance Testing

EXPERIMENT. Environmental Influences on Microbial Growth Salt Tolerance Testing EXPERIMENT Environmental Influences on Microbial Growth Salt Tolerance Testing Hands-On Labs, Inc. Version 42-0307-00-01 Review the safety materials and wear goggles when working with chemicals. Read the

More information

BACTERIAL GENETICS: Labs I & II

BACTERIAL GENETICS: Labs I & II BACTERIAL GENETICS: Labs I & II The Bacterial Genetics Labs will extend over two laboratory periods. During the first lab, you will set up two different experiments using the bacterium Escherichia coli.

More information

Standard Operating Procedure Title: Good laboratory practices (GLP) for microbiology and chemistry laboratories

Standard Operating Procedure Title: Good laboratory practices (GLP) for microbiology and chemistry laboratories Standard Operating Procedure Title: Good laboratory practices (GLP) for microbiology and chemistry laboratories Department Micro Laboratory Document no MICLAB 155 Title Good laboratory practices (GLP)

More information

Microbial assay measures the activity of antibiotics (Extent of ability to inhibit

Microbial assay measures the activity of antibiotics (Extent of ability to inhibit 6. MICROBIOLOGICAL EVALUATION Microbial assay measures the activity of antibiotics (Extent of ability to inhibit the growth of micro organism) or vitamins and amino acids (Extent to support the growth

More information

TRYPTIC SOY AGAR (TSA) WITH LECITHIN AND TWEEN 80

TRYPTIC SOY AGAR (TSA) WITH LECITHIN AND TWEEN 80 TRYPTIC SOY AGAR (TSA) WITH LECITHIN AND TWEEN 80 Cat. no. P45 TSA with Lecithin and Tween 80, 15x60mm Contact Plate, 15ml Cat. no. Q13 TSA with Lecithin and Tween 80, 20x125mm Tube, 18ml Deep Cat. no.

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

Principle of Lab. Safety

Principle of Lab. Safety Sulaimani University College of Pharmacy Principle of Lab. Safety Dr. Abdullah Ahmed Hama 1 1. Wear appropriate clothing and shoes to the laboratory. Shoes must completely cover the feet to provide protection

More information

Standard Operating Procedure Title: Stock Suspensions of Micro-Organisms

Standard Operating Procedure Title: Stock Suspensions of Micro-Organisms 2. Correct Aseptic technique must be used when performing all Microbiological procedures. 3. All work must be carried out in the Biohazard Cabinet. Table of Contents 2. Positive Identification of Reference

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

Document No. FTTS-FA-001. Specified Requirements of Antibacterial Textiles for General Use

Document No. FTTS-FA-001. Specified Requirements of Antibacterial Textiles for General Use 1. Purpose and Scope This criterion is applicable to the evaluation and testing of antibacterial activity of textile for general use. The quantitative evaluation of antibacterial activity is judged by

More information

Bacterial Counts - Quantitative Analysis of Microbes

Bacterial Counts - Quantitative Analysis of Microbes Bacterial Counts - Quantitative Analysis of Microbes Introduction: It is often important to know not only what types of bacteria are in a sample but also how many of them are present. Food manufacturers

More information

CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS

CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS Specific control measures can be used to kill or inhibit the growth of microorganisms. A procedure which leads to the death of cells is broadly

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

Test Method of Specified Requirements of Antibacterial Textiles for Medical Use FTTS-FA-002

Test Method of Specified Requirements of Antibacterial Textiles for Medical Use FTTS-FA-002 Test Method of Specified Requirements of Antibacterial Textiles for Medical Use FTTS-FA-002 FTTS-FA-002 Antibacterial Textiles for Medical Use Antibacterial Textiles suppress and even kill harmful bacteria

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

Test Method for the Continuous Reduction of Bacterial Contamination on Copper Alloy Surfaces

Test Method for the Continuous Reduction of Bacterial Contamination on Copper Alloy Surfaces Test Method for the Continuous Reduction of Bacterial Contamination on Copper Alloy Surfaces Test Organisms: Staphylococcus aureus (ATCC 6538) Enterobacter aerogenes (ATCC 13048) Pseudomonas aeruginosa

More information

ANALYTICAL REPORT: Comparison of the Microbial Recovery Efficacy of QI Medical EnviroTest Paddles versus a Conventional Contact Plate

ANALYTICAL REPORT: Comparison of the Microbial Recovery Efficacy of QI Medical EnviroTest Paddles versus a Conventional Contact Plate FOCUS Scientific Services LLC ANALYTICAL REPORT: Comparison of the Microbial Recovery Efficacy of QI Medical EnviroTest Paddles versus a Conventional Contact Plate REPORT: FS-QI-GM-003 Study Manager: Anthony

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

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

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

Bacteria and other microbes have particular requirements for growth When they reside in and on our bodies or in the environment, they harvest their

Bacteria and other microbes have particular requirements for growth When they reside in and on our bodies or in the environment, they harvest their Bacteria and other microbes have particular requirements for growth When they reside in and on our bodies or in the environment, they harvest their food from us or from the environment When we grow bacteria

More information

I January 23-January 26 INTRODUCTION; SAFETY; ASEPTIC TECHNIQUE; USE OF MICROSCOPES; OBSERVATION OF PREPARED SLIDES; ENVIRONMENTAL SAMPLE; EPIDEMIC

I January 23-January 26 INTRODUCTION; SAFETY; ASEPTIC TECHNIQUE; USE OF MICROSCOPES; OBSERVATION OF PREPARED SLIDES; ENVIRONMENTAL SAMPLE; EPIDEMIC MICROBIOLOGY LABORATORY (BIOL 310L) SCHEDULE Spring 2017 Lecture Professor: Dr. Susan Morrison Lab Instructors: Ms. Tracy Hirsch Dr. Susan Morrison Required: (1) Leboffe & Pierce, Microbiology Laboratory

More information

Test Method for Efficacy of Copper Alloy Surfaces as a Sanitizer

Test Method for Efficacy of Copper Alloy Surfaces as a Sanitizer Test Method for Efficacy of Copper Alloy Surfaces as a Sanitizer Test Organisms: Staphylococcus aureus (ATCC 6538) Enterobacter aerogenes (ATCC 13048) Pseudomonas aeruginosa (ATCC 15442) Methicillin Resistant

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

Presto 96 Well gdna Bacteria Kit

Presto 96 Well gdna Bacteria Kit Presto 96 Well gdna Bacteria Kit 96GBB02 (2 x 96 well plates/kit) 96GBB04 (4 x 96 well plates/kit) 96GBB10 (10 x 96 well plates/kit) Instruction Manual Ver. 05.04.17 For Research Use Only Advantages Sample:

More information

Large Volume Serial Dilutions:

Large Volume Serial Dilutions: Serial Dilutions All three bacterial plate count methods described in lab require you to serially dilute your samples until you have 30-300 colony forming units (CFU) on the plate. Plates with more than

More information

CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS

CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS CONTROL OF MICROBIAL GROWTH - DISINFECTANTS AND ANTISEPTICS Specific control measures can be used to kill or inhibit the growth of microorganisms. A procedure which leads to the death of cells is broadly

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

Project 5: Urine Cultures and Identification

Project 5: Urine Cultures and Identification Project 5: Urine Cultures and Identification Readings: http://www.webmd.com/a-to-z-guides/urine-culture http://www.medscape.com/viewarticle/558845 (Listen to the two lectures by Dr. Robert A. Weinstein.)

More information

Adapted from Biology 15 Laboratory Manual Supplement: Wrightsman, Ininns and Cannon-Moloznic, Saddleback College, CA 92692

Adapted from Biology 15 Laboratory Manual Supplement: Wrightsman, Ininns and Cannon-Moloznic, Saddleback College, CA 92692 Biology 4B Laboratory Bacteriological Examination of Water Adapted from Biology 15 Laboratory Manual Supplement: Wrightsman, Ininns and Cannon-Moloznic, Saddleback College, CA 92692 Objectives Carry out

More information

UNIVERSITEIT GENT. Laboratory of Microbiology K.L. Ledeganckstr. 35 B-9000 Gent (BELGIUM) SOP. Standard Operating Procedure.

UNIVERSITEIT GENT. Laboratory of Microbiology K.L. Ledeganckstr. 35 B-9000 Gent (BELGIUM) SOP. Standard Operating Procedure. SOP Standard Operating Procedure Author: Acronym: Date last modified: Geert Huys ASIARESIST-PRES 20-11-2002 Title: PRESERVATION OF BACTERIA USING COMMERCIAL CRYOPRESERVATION SYSTEMS References: Reviewed

More information

Actero Salmonella/STEC Enrichment Media Product Information

Actero Salmonella/STEC Enrichment Media Product Information Actero Salmonella/STEC Enrichment Media Product Information INTENDED USE: Actero Salmonella/STEC Enrichment Media is a selective medium optimized for an improved enrichment of Salmonella spp. from food

More information

Bio 205 & Bio 298AA Fall 2015 Laboratory Policies and Schedule

Bio 205 & Bio 298AA Fall 2015 Laboratory Policies and Schedule Bio 205 & Bio 298AA Fall 2015 Laboratory Policies and Schedule It is your responsibility to be familiar with the policies given in this syllabus. Prof. Anne Healy Office phone: 480-423-6779 E-mail: anne.cedergren-healy@sccmail.maricopa.edu

More information

MICROBIOLOGICAL TOOLS FOR QUALITY ASSURANCE IN HATCHERY: Laboratory Methods

MICROBIOLOGICAL TOOLS FOR QUALITY ASSURANCE IN HATCHERY: Laboratory Methods Issue No.29 / March 2010 MICROBIOLOGICAL TOOLS FOR QUALITY ASSURANCE IN HATCHERY: Laboratory Methods By Dr Vincent TURBLIN, Deputy Regional Market Manager Poultry - CEVA Animal Health Asia Pacific Most

More information

Aerobic Count. Interpretation Guide. 3M Food Safety 3M Petrifilm Aerobic Count Plate

Aerobic Count. Interpretation Guide. 3M Food Safety 3M Petrifilm Aerobic Count Plate 3M Food Safety 3M Petrifilm Aerobic Count Plate Aerobic Count Interpretation Guide The 3M Petrifilm Aerobic Count (AC) Plate is a ready-made culture medium system that contains Standard Methods nutrients,

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

Inoculate: Media. Physical State of Media: Liquid. The Five I s: Basic Techniques to Culture Microbes Tools of the Microbiology Laboratory

Inoculate: Media. Physical State of Media: Liquid. The Five I s: Basic Techniques to Culture Microbes Tools of the Microbiology Laboratory The Five I s: Basic Techniques to Culture Microbes Tools of the Microbiology Laboratory 1. Inoculate 2. Incubate 3. Isolate 4. Inspect 5. Identify The Five I s: Inoculate Inoculate: Media Classified according

More information

Culturing microorganisms

Culturing microorganisms Culturing microorganisms I. Historical development II. Problems and Solutions III. Studying microorganisms without a microscope -- culturing techniques A. How do you do it? B. Inoculation and isolation

More information

BIOLOGY 163 LABORATORY. THE EFFECT OF ANTIBIOTICS ON THE GROWTH OF Escherichia coli B (Revised Fall 2014)

BIOLOGY 163 LABORATORY. THE EFFECT OF ANTIBIOTICS ON THE GROWTH OF Escherichia coli B (Revised Fall 2014) BIOLOGY 163 LABORATORY THE EFFECT OF ANTIBIOTICS ON THE GROWTH OF Escherichia coli B (Revised Fall 2014) Bacteria are single-celled prokaryotic organisms. As bacterial cells take in nutrients from their

More information

Actero Salmonella Enrichment Media Product Information

Actero Salmonella Enrichment Media Product Information Actero Salmonella Enrichment Media Product Information INTENDED USE: Actero Salmonella Enrichment Media is a selective medium optimized for an improved enrichment of Salmonella spp. from food and environmental

More information

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 Division of Environmental Health Sciences Albany, New York NYS DOH LEB-606 Identification

More information

Culturing microorganisms

Culturing microorganisms Culturing microorganisms I. Historical development II. Problems and Solutions III. Studying microorganisms without a microscope -- culturing techniques A. How do you do it? B. Inoculation and isolation

More information

Ch 6. Microbial Growth

Ch 6. Microbial Growth Ch 6 Microbial Growth Student Learning Outcomes: Classify microbes into five groups on the basis of preferred temperature range. Explain the importance of osmotic pressure to microbial growth. Provide

More information

Standard Operating Procedure Safe Autoclave Operations

Standard Operating Procedure Safe Autoclave Operations Standard Operating Procedure Safe Autoclave Operations The purpose of this instructional document is to introduce and familiarize the reader to the standard operating procedures for the safe use of autoclaves.

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

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

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765

New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 New York State Department of Health - Wadsworth Center Laboratory of Environmental Biology NYS ELAP Laboratory ID 10765 Division of Environmental Health Sciences Albany, New York NYS DOH LEB-606 Identification

More information