MicroStation System, MicroLog Version 4.2 - DTU Systems Biology ...

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Preparing Samples Characterizing Anaerobic Bacteria Gram-negative anaerobic rods require further investigation to determine proper handling and identification. Characterizing rapidly-growing, gram-negative microbes If your microbe is rod-shaped, anaerobic, gram negative, and grows rapidly, a kanamycin disk test will help differentiate Fusobacterium from Bacteroides/Prevotella. This has important implications in regards to choosing the appropriate inoculation protocol. Performing a kanamycin disk test • Streak rapidly-growing, gram-negative rods on BUA plates. • Add a 1 mg kanamycin disk to the first quadrant (immediately after streaking, before incubating the organism). • Incubate the plate overnight. • If the bacterium is sensitive (a 10 mm zone of clearing will surround the kanamycin disk), you have a presumptive Fusobacterium. • If the bacterium is resistant (no or
Preparing Samples Media shorthand: B = blood M = maltose T = thioglycolate ME = malt extract Culturing Your Microbe From a single isolated colony subculture your microbe onto the proper agar medium. Most of the species in Biolog’s database are relatively fast growers. However, if your microbe grows slowly, you may need to streak (subculture) more than one agar plate. Incubate according to instructions in package insert. In general: • Incubate most organisms for 16-24 hours. Very slow-growing species and yeasts may require 48 hours. Slow-growing anaerobic bacteria may require even longer incubation. Incubate filamentous fungi for 5-10 days (sporulation of isolate). Slower growing filamentous fungi may require longer incubation for sporulation. • Don’t over-incubate your culture. Excess incubation can cause microbes to enter a stationary phase, during which they lose viability and metabolic activity. • For best results, subculture anaerobic bacteria twice on BUA. Prepare the suspension from the second plate only. • For filamentous fungi, subculture less hyphae for fast growing fungi and more hyphae for slow growing fungi. Table 4-1 and Appendix 2 will help you select the correct culture medium. Organism Type Organism Abbreviations Gram Negative Non-Enteric** GN-NENT BUG + B Gram Negative Enteric GN-ENT BUG + B Gram Negative Fastidious GN-FAS Chocolate Gram Positive Cocci Gram Positive Rods (non-spore forming) Gram Positive Rods (spore forming bacillus) GP-COCCUS GP-ROD BUG + B Culture Media GP-ROD SB BUG + M + T Anaerobes AN BUA + B Yeasts YT BUY Filamentous Fungi (and select yeast species) FF 2% ME TABLE 4-1: SELECTING THE CORRECT CULTURE MEDIUM **Many agriculture organisms should be grown on BUG (w/o Blood). For listing, see Appendix 4. Section 4 � Page 6 MicroStation System/MicroLog User Guide Apr 07
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Page 3 and 4: LICENSE AGREEMENT Notice: This is a
Page 5 and 6: Table of Contents Section Title 1.
Page 7 and 8: 10. Frequently Asked Questions…
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Page 15 and 16: 8. E-mail the User Key File to tech
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Page 19 and 20: Follow directions closely to obtain
Page 29 and 30: Preparing Samples Preparing an inoc
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Page 47 and 48: Reading MicroPlates Set Up tab Sele
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Page 61 and 62: Reading MicroPlates Using Worksheet
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Page 69 and 70: Interpreting Results Understanding
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Data Management and Compiling Datab
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Combining Data Files Data Managemen
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OK, Out, Hold, Atypical Sorting lab
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Remember! When building your own da
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DATABASE WINDOW Data Management and
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Arrow indicates selected entry Data
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Export Help Mark this choice if you
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8. Administration and Security In t
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Important! • If you test Password
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Log-In Privileges Administration an
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Remember that the password must be
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9. Technical Notes In this section:
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Making Biolog Universal Growth Agar
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� � � � Technical Notes Gen
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10. Frequently Asked Questions In t
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Q A Q A Q A Frequently Asked Questi
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Troubleshooting Symptom Cause Solut
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Troubleshooting Preparing Inocula S
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Troubleshooting MicroStation Reader
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Glossary Dendrogram Cluster diagram
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Glossary Spore, fungal Sterile, fun
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Culture Media Atmosphere Temperatur
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Appendices: Notes Notes: Section 13
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Appendix 4: Database Species Lists
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Appendix 5: Program Printouts Appen
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Appendix 5: Program Printouts 1�
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Appendix 5: Program Printouts Funga
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Appendix 5: Program Printouts Print
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Appendix 6: Dangerous Pathogens (DP
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MicroStation System, MicroLog Version 4.2 - DTU Systems Biology ...

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