Maintenance of Bacterial Strains
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Exploring Small-Scale Chemostats to Scale up Microbial Processes: 3-Hydroxypropionic Acid Production in S
Lawrence Berkeley National Laboratory Recent Work Title Exploring small-scale chemostats to scale up microbial processes: 3-hydroxypropionic acid production in S. cerevisiae. Permalink https://escholarship.org/uc/item/43h8866j Journal Microbial cell factories, 18(1) ISSN 1475-2859 Authors Lis, Alicia V Schneider, Konstantin Weber, Jost et al. Publication Date 2019-03-11 DOI 10.1186/s12934-019-1101-5 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Lis et al. Microb Cell Fact (2019) 18:50 https://doi.org/10.1186/s12934-019-1101-5 Microbial Cell Factories RESEARCH Open Access Exploring small-scale chemostats to scale up microbial processes: 3-hydroxypropionic acid production in S. cerevisiae Alicia V. Lis1, Konstantin Schneider1,2, Jost Weber1,3, Jay D. Keasling1,4,5,6,7, Michael Krogh Jensen1 and Tobias Klein1,2* Abstract Background: The physiological characterization of microorganisms provides valuable information for bioprocess development. Chemostat cultivations are a powerful tool for this purpose, as they allow defned changes to one single parameter at a time, which is most commonly the growth rate. The subsequent establishment of a steady state then permits constant variables enabling the acquisition of reproducible data sets for comparing microbial perfor- mance under diferent conditions. We performed physiological characterizations of a 3-hydroxypropionic acid (3-HP) producing Saccharomyces cerevisiae strain in a miniaturized and parallelized chemostat cultivation system. The physi- ological conditions under investigation were various growth rates controlled by diferent nutrient limitations (C, N, P). Based on the cultivation parameters obtained subsequent fed-batch cultivations were designed. Results: We report technical advancements of a small-scale chemostat cultivation system and its applicability for reliable strain screening under diferent physiological conditions, i.e. -
Detection of Acid-Producing Bacteria Nachweis Von Säureproduzierenden Bakterien Détection De Bactéries Produisant Des Acides
(19) TZZ ¥ _T (11) EP 2 443 249 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12Q 1/04 (2006.01) G01N 33/84 (2006.01) 19.11.2014 Bulletin 2014/47 (86) International application number: (21) Application number: 10790013.6 PCT/US2010/038569 (22) Date of filing: 15.06.2010 (87) International publication number: WO 2010/147918 (23.12.2010 Gazette 2010/51) (54) DETECTION OF ACID-PRODUCING BACTERIA NACHWEIS VON SÄUREPRODUZIERENDEN BAKTERIEN DÉTECTION DE BACTÉRIES PRODUISANT DES ACIDES (84) Designated Contracting States: (74) Representative: Isarpatent AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Patent- und Rechtsanwälte GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO Friedrichstrasse 31 PL PT RO SE SI SK SM TR 80801 München (DE) (30) Priority: 15.06.2009 US 187107 P (56) References cited: 15.03.2010 US 314140 P US-A- 4 528 269 US-A- 5 098 832 US-A- 5 164 301 US-A- 5 601 998 (43) Date of publication of application: US-A- 5 601 998 US-A- 5 786 167 25.04.2012 Bulletin 2012/17 US-B2- 6 756 225 US-B2- 7 150 977 (73) Proprietor: 3M Innovative Properties Company • DARUKARADHYA J ET AL: "Selective Saint Paul, MN 55133-3427 (US) enumeration of Lactobacillus acidophilus, Bifidobacterium spp., starter lactic acid bacteria (72) Inventors: and non-starter lactic acid bacteria from Cheddar • YOUNG, Robert, F. cheese", INTERNATIONAL DAIRY JOURNAL, Saint Paul, Minnesota 55133-3427 (US) ELSEVIER APPLIED SCIENCE, BARKING, GB, • MACH, Patrick, A. -
Chemostat Culture for Yeast Experimental Evolution
Downloaded from http://cshprotocols.cshlp.org/ at Cold Spring Harbor Laboratory Library on August 9, 2017 - Published by Cold Spring Harbor Laboratory Press Protocol Chemostat Culture for Yeast Experimental Evolution Celia Payen and Maitreya J. Dunham1 Department of Genome Sciences, University of Washington, Seattle, Washington 98195 Experimental evolution is one approach used to address a broad range of questions related to evolution and adaptation to strong selection pressures. Experimental evolution of diverse microbial and viral systems has routinely been used to study new traits and behaviors and also to dissect mechanisms of rapid evolution. This protocol describes the practical aspects of experimental evolution with yeast grown in chemostats, including the setup of the experiment and sampling methods as well as best laboratory and record-keeping practices. MATERIALS It is essential that you consult the appropriate Material Safety Data Sheets and your institution’s Environmental Health and Safety Office for proper handling of equipment and hazardous material used in this protocol. Reagents Defined minimal medium appropriate for the experiment For examples, see Protocol: Assembly of a Mini-Chemostat Array (Miller et al. 2015). Ethanol (95%) Glycerol (20% and 50%; sterile) Yeast strain of interest Equipment Agar plates (appropriate for chosen strain) Chemostat array Assemble the apparatus as described in Miller et al. (2013) and Protocol: Assembly of a Mini-Chemostat Array (Miller et al. 2015). Cryo deep-freeze labels Cryogenic vials Culture tubes Cytometer (BD Accuri C6) Glass beads, 4 mm (sterile; for plating yeast cells) Glass cylinder Kimwipes 1Correspondence: [email protected] © 2017 Cold Spring Harbor Laboratory Press Cite this protocol as Cold Spring Harb Protoc; doi:10.1101/pdb.prot089011 559 Downloaded from http://cshprotocols.cshlp.org/ at Cold Spring Harbor Laboratory Library on August 9, 2017 - Published by Cold Spring Harbor Laboratory Press C. -
Microbiology Laboratory Exercises Third Edition 2020
MICROBIOLOGY Laboratory Exercises Third Edition Keddis & Rauschenbach 2020 Photo Credits (in order of contribution): Diane Davis, Ines Rauschenbach & Ramaydalis Keddis Acknowledgements: Many thanks to those in the Department of Biochemistry and Microbiology, Rutgers University, who have through the years inspired our enthusiasm for the science and teaching of microbiology, with special thanks to Diane Davis, Douglas Eveleigh and Max Häggblom. Safety: The experiments included in this manual have been deemed safe by the authors when all necessary safety precautions are met. The authors recommend maintaining biosafety level 2 in the laboratory setting and using risk level 1 organisms for all exercises. License: This work is licensed under a Creative Commons Attribution- NonCommercial-NoDerivatives 4.0 International License Microbiology Laboratory Exercises Third Edition 2020 Ramaydalis Keddis, Ph.D. Ines Rauschenbach, Ph.D. Department of Biochemistry and Microbiology Rutgers, The State University of New Jersey CONTENTS PAGE Introduction Schedule ii Best Laboratory Practices Iii Working in a Microbiology Laboratory iv Exercises Preparation of a Culture Medium 1 Culturing and Handling Microorganisms 3 Isolation of a Pure Culture 5 Counting Bacterial Populations 8 Controlling Microorganisms 10 Disinfectants 10 Antimicrobial Agents: Susceptibility Testing 12 Hand Washing 14 The Lethal Effects of Ultraviolet Light 15 Selection of Fungi from Air 17 Microscopy 21 Morphology and Staining of Bacteria 26 Microbial Metabolism 30 Enzyme Assay 32 Metabolic -
Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation
City University of New York (CUNY) CUNY Academic Works Open Educational Resources Queensborough Community College 2016 Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation Joan Petersen CUNY Queensborough Community College Susan McLaughlin CUNY Queensborough Community College How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qb_oers/16 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation By Dr. Susan McLaughlin & Dr. Joan Petersen Queensborough Community College Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation Table of Contents Preface………………………………………………………………………………………i Acknowledgments…………………………………………………………………………..ii Microbiology Lab Safety Instructions…………………………………………………...... iii Lab 1. Introduction to Microscopy and Diversity of Cell Types……………………......... 1 Lab 2. Introduction to Aseptic Techniques and Growth Media………………………...... 19 Lab 3. Preparation of Bacterial Smears and Introduction to Staining…………………...... 37 Lab 4. Acid fast and Endospore Staining……………………………………………......... 49 Lab 5. Metabolic Activities of Bacteria…………………………………………….…....... 59 Lab 6. Dichotomous Keys……………………………………………………………......... 77 Lab 7. The Effect of Physical Factors on Microbial Growth……………………………... 85 Lab 8. Chemical Control of Microbial Growth—Disinfectants and Antibiotics…………. 99 Lab 9. The Microbiology of Milk and Food………………………………………………. 111 Lab 10. The Eukaryotes………………………………………………………………........ 123 Lab 11. Clinical Microbiology I; Anaerobic pathogens; Vectors of Infectious Disease….. 141 Lab 12. Clinical Microbiology II—Immunology and the Biolog System………………… 153 Lab 13. Putting it all Together: Case Studies in Microbiology…………………………… 163 Appendix I. -
Laboratory Equipment Reference Sheet
Laboratory Equipment Stirring Rod: Reference Sheet: Iron Ring: Description: Glass rod. Uses: To stir combinations; To use in pouring liquids. Evaporating Dish: Description: Iron ring with a screw fastener; Several Sizes Uses: To fasten to the ring stand as a support for an apparatus Description: Porcelain dish. Buret Clamp/Test Tube Clamp: Uses: As a container for small amounts of liquids being evaporated. Glass Plate: Description: Metal clamp with a screw fastener, swivel and lock nut, adjusting screw, and a curved clamp. Uses: To hold an apparatus; May be fastened to a ring stand. Mortar and Pestle: Description: Thick glass. Uses: Many uses; Should not be heated Description: Heavy porcelain dish with a grinder. Watch Glass: Uses: To grind chemicals to a powder. Spatula: Description: Curved glass. Uses: May be used as a beaker cover; May be used in evaporating very small amounts of Description: Made of metal or porcelain. liquid. Uses: To transfer solid chemicals in weighing. Funnel: Triangular File: Description: Metal file with three cutting edges. Uses: To scratch glass or file. Rubber Connector: Description: Glass or plastic. Uses: To hold filter paper; May be used in pouring Description: Short length of tubing. Medicine Dropper: Uses: To connect parts of an apparatus. Pinch Clamp: Description: Glass tip with a rubber bulb. Uses: To transfer small amounts of liquid. Forceps: Description: Metal clamp with finger grips. Uses: To clamp a rubber connector. Test Tube Rack: Description: Metal Uses: To pick up or hold small objects. Beaker: Description: Rack; May be wood, metal, or plastic. Uses: To hold test tubes in an upright position. -
Laboratory Supplies and Equipment
Laboratory Supplies and Equipment Beakers: 9 - 12 • Beakers with Handles • Printed Square Ratio Beakers • Griffin Style Molded Beakers • Tapered PP, PMP & PTFE Beakers • Heatable PTFE Beakers Bottles: 17 - 32 • Plastic Laboratory Bottles • Rectangular & Square Bottles Heatable PTFE Beakers Page 12 • Tamper Evident Plastic Bottles • Concertina Collapsible Bottle • Plastic Dispensing Bottles NEW Straight-Side Containers • Plastic Wash Bottles PETE with White PP Closures • PTFE Bottle Pourers Page 39 Containers: 38 - 42 • Screw Cap Plastic Jars & Containers • Snap Cap Plastic Jars & Containers • Hinged Lid Plastic Containers • Dispensing Plastic Containers • Graduated Plastic Containers • Disposable Plastic Containers Cylinders: 45 - 48 • Clear Plastic Cylinder, PMP • Translucent Plastic Cylinder, PP • Short Form Plastic Cylinder, PP • Four Liter Plastic Cylinder, PP NEW Polycarbonate Graduated Bottles with PP Closures Page 21 • Certified Plastic Cylinder, PMP • Hydrometer Jar, PP • Conical Shape Plastic Cylinder, PP Disposal Boxes: 54 - 55 • Bio-bin Waste Disposal Containers • Glass Disposal Boxes • Burn-upTM Bins • Plastic Recycling Boxes • Non-Hazardous Disposal Boxes Printed Cylinders Page 47 Drying Racks: 55 - 56 • Kartell Plastic Drying Rack, High Impact PS • Dynalon Mega-Peg Plastic Drying Rack • Azlon Epoxy Coated Drying Rack • Plastic Draining Baskets • Custom Size Drying Racks Available Burn-upTM Bins Page 54 Dynalon® Labware Table of Contents and Introduction ® Dynalon Labware, a leading wholesaler of plastic lab supplies throughout -
ED149972.Pdf
-74 ..t.rcrVtr.,,,.. r . ", 0 ' 4^Az7 .. ' . DOCUMNIT ant= 2D 149 972 SE 023- 413 AUTHOR . We'sterhold, Arnold F.., Ed.; 'Bennett; Ernest C., . Ed. , _ TITLE : Ahanual of Simplified Laboratory Methods for . Operators of 'Wastewater Treatment Facilities.. INSTITUTION' Illinois State Epvircidental.Protection Agencyc , .. - 'Springfield.. PUB DATE Apr 74 NOTE 94p.; Pages 1-1 through 1-12 (General Introduction) removed due to copyright restrictions; Section 8 'missing; Contains occasional light type; Best Copy / Available EDRS PRICE .M7-$0.83 HC-$4.67 Plus Postage. DESCRIPTORS, Chemistry; Environmental Education; *Environmental Technicians; Independent Study; *Instructional Materials; Job Skills; *Laboratory Tchniques; *Pollution; *Post Seccndary Education; Public Health; *Water Pollution Control IDENTIFIERS cf*Waste Water Treatment ABSTRACT . ./ This manual is designed to provide the seall wastewater treatment plant operator, as well as the new or inexperienced operator, with simplified methods for laboratory analysis of'water"and wastewater. It is emphasized that this manual is not a replacement for standard methods but a guide for plantswith insufficient equipment to pefform analyses ifi..accordance with ,standard methods. Eacj, of the sections is designed to becomplete I within itself. The tests and measurements 'presented include: acids, biochethical oxygen demand (BOD);40674plved oxygen, residues, sludge, a'nd suspended solids. (CS) el. -V or 4. ***********44*************************************i*******i*******4S*** I4 * Reproductions -
Method 1604: Total Coliforms and Escherichia Coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (MI Medium)
Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (MI Medium) September 2002 U.S. Environmental Protection Agency Office of Water (4303T) 1200 Pennsylvania Avenue, NW Washington, DC 20460 EPA-821-R-02-024 Disclaimer The Engineering and Analysis Division, of the Office of Science and Technology, has reviewed and approved this report for publication. The Office of Science and Technology directed, managed, and reviewed the work of DynCorp in preparing this report. Neither the United States Government nor any of its employees, contractors, or their employees make any warranty, expressed or implied, or assumes any legal liability or responsibility for any third party’s use of or the results of such use of any information, apparatus, product, or process discussed in this report, or represents that its use by such party would not infringe on privately owned rights. The content of this method version is identical to the February 2000 version of Membrane Filter Method for the Simultaneous Detection of Total Coliforms and Escherichia coli in Drinking Water (EPA-600-R- 00-013) with one exception, the addition of MI broth. Since MI broth was approved on November 6, 2001, as a minor modification of the MI agar method, it has also been included in this document. Mention of trade names or commercial products does not constitute endorsement or recommendation for use. Questions concerning this method or its application should be addressed to: Robin K. Oshiro Engineering and Analysis Division (4303T) U.S. EPA Office of Water, Office of Science and Technology 1200 Pennsylvania Avenue, NW Washington, DC 20460 [email protected] 202-566-1075 202-566-1053 (facsimile) Table of Contents 1.0 Scopeand Application ........................................................ -
Qualitube™ Kit for Microcystin
™ QualiTube Kit For Microcystin Highlights: Catalog Number ET 022 Semi-quantitative field screening Intended Use of Microcystin toxin in surface water The EnviroLogix QualiTube Kit for Microcystin is designed for semi- Detects from 0.5 to 3 ppb quantitative field screening of Microcystin toxin in surface water samples. The kit is supplied with calibrators at 0.5 and 3 ppb. The assay range can easily be extended. How the Test Works Contents of Kit: The QualiTube Kit for Microcystin is a competitive Enzyme-Linked ImmunoSorbent Assay (ELISA). In the test, Microcystin toxin in the sample 36 antibody-coated test tubes competes with enzyme (horseradish peroxidase)-labeled Microcystin for a 1 vial of 0.5 ppb Microcystin LR limited number of antibody binding sites on the inside surface of the test Calibrator tubes. 1 vial of 3 ppb Microcystin LR Calibrator After a simple wash step, the outcome of the competition is visualized with 1 dropper bottle of Assay Diluent a color development step. As with all competitive immunoassays, sample 1 dropper bottle of Microcystin- concentration is inversely proportional to color development. enzyme Conjugate 1 dropper bottle of Substrate Darker color = Lower concentration 1 bottle of Stop Solution Lighter color = Higher concentration 36 sample pipettes Limit of Detection The Limit of Detection (LOD) of the EnviroLogix Microcystin Tube Kit is Optional Accessory Item: 0.3 ppb. The LOD was determined by interpolation at 81.5% B0* from a ACC 062 – 1 vial of 1.5 ppb standard curve. 81.5% B0 was determined to be 2 standard deviations from Microcystin LR Calibrator the mean of a population of negative water samples. -
Hitouch™ E.Coli/ Coliform Count Flexi Plate FL002
HiTouch™ E.Coli/ Coliform Count Flexi Plate FL002 For enumeration (count) of all coliform along with differential count of E.coli. Composition** Ingredients Gms / Litre Tryptose 14.000 Peptone special 5.000 Bile salts mixture 1.500 Disodium hydrogen phosphate 1.000 Monosodium dihydrogen phosphate 0.600 Sodium chloride 2.400 Chromogen 0.075 Indicator dye 0.100 Agar 12.000 Final pH ( at 25°C) 7.3±0.2 **Formula adjusted, standardized to suit performance parameters Directions Open the pouch in the protected area . Remove the wrappping and open the lid and carefully lift up the enclosed prepared medium plate so as to avoid touching the agar surface by hand.Touch the surface of agar plate onto the surface to be tested. Gently press the plate manually for upto 10 sencond. Apply constant and uniform pressure to the whole surface (ensuring that an even pressure of 25 gm/cm2 is distributed over the whole plate for 10 seconds). Replace exposed medium plate back in the base plate. Close the lid. Press the sides of the lid to make sure that it is fixed in the grooves. Disinfect the surface where the sample was taken in order to remove any possible traces of agar. Incubate the plates at specified temperature. After incubation as recommended count the number of colonies which have appeared on the surface of medium. Alternative Methods of Inoculation : To use as Culture Plate (ii), Sample Dilution Plate (iii) or Swabbing Plate (iv) To use as Gravitation Settling Plate (v).These plates are specially developed for microbial testing,The Flexi plate medium formula is suitable for enumeration ofyeast and mould(fungal) population and the grids enable direct reading on the plates of the number of colonies per cm2. -
Agar Plate Storage in the Fridge
Published on ASSIST (https://assist.asta.edu.au) Home > Agar plate storage in the fridge Agar plate storage in the fridge Posted by Anonymous on Thu, 2018-09-06 16:31 Agar plate storage in the fridge: Is it necessary to store the agar plates upside down in the fridge? As in the school we are attending to so many things that we are sometimes in a hurry to check if all the agar plates have gelled but stack and seal and put them in fridge for further use. Yes we placing definitely siting them in the incubator upside down so that condensation doesn't fall on the agar plates. Another question is how long can we store the agar plates in the fridge? Voting: 0 No votes yet Year Level: 9 10 Senior Secondary Laboratory Technicians: Laboratory Technicians Showing 1-1 of 1 Responses Agar plate storage in the fridge Submitted by sat on 06 September 2018 Correct aseptic preparation and storage of agar plates away from light and heat is necessary to protect them from contamination, dehydration and degradation of chemical constituents. Setting of agar plates: Agar plates when poured should be allowed to set undisturbed at room temperature. It is best practice that when completely set and cooled to room temperature that they should be stored stacked upside down in sealed plastic bags in the fridge at 4 °C. It is a good idea to reuse the plastic bags from which the sterile Petri dishes came from for storage. Storage this way will prevent: Condensation that develops on the lid from dripping onto the agar surface which is a potential source of contamination.