Navigation Links
Isolation of Microorganisms

Jrgen Frhlich, Helmut Knig and Dietmar Kahle
Jrgen Frhlich and Helmut Knig; Institute for Microbiology and Wine Research,
Johannes-Gutenberg-University; Mainz, Germany
Dietmar Kahle, Eppendorf AG Problem The isolation of microorganisms (such as bacteria or yeast) from complex mixed cultures and their cultivation in a pure culture is an essential prerequisite for their precise identification and characterization.

Several different processes are used for this purpose, including the classic platecasting process invented by Robert Koch, as well as dilution methods or selective enrichment.

However, it can prove to be very difficult to isolate microbes when the conditions for their growth are not known or when the microbes grow in a heterogeneous mixed culture in very small numbers only.

Such experiments are now possible thanks to new techniques which have been developed recently (e.g., optical laser tweezers).

This article outlines a novel method which, with the aid of a micromanipulation workstation, enables the isolation of individual bacteria from a mixed culture.

These cells can then be cultivated further or analyzed via single cell PCR*.

Isolation of individual bacteria Material The workstation consists of an inverse research microscope with up to 100-fold magnification (phase contrast), an Eppendorf Micromanipulator 5171 or TransferMan device as well as the Eppendorf CellTram Oil hydraulic piston pump.

Isolation was carried out with special sterile glass capillaries. The dimensions of these microcapillaries depend on the size of the cells to be isolated. For typical experiments, capillaries with an inner diameter of 5 m and a tip angle of 45 were used. These capillaries were designed in accordance with our specifications (Eppendorf).

Dried bacteria cultures were resuspended using suitable sterile nutrient solutions. The isolated cells were then transferred into the appropriate micro test tubes for further processing.

Advantages
  • This process can be used for cloning and isolating microorganisms (such as bacteria, archaea, protozoa and fungi) as well as eukaryotic organelles and cell nuclei.
  • The risk of contamination during the identification of non-cultivatable microorganisms from mixed cultures by means of single cell PCR is greatly reduced.
  • Microorganisms present in low numbers can also be examined.
  • In addition to aerobic microorganisms, it is also possible to isolate anaerobic microorganisms (with the appropriate accessories).
  • The electronic control of the micro-manipulator and the high-resolution piston pump guarantee rapid and simple isolation.
Conclusion Microorganisms which were previously difficult to access can now be isolated and characterized rapidly using this simple and relatively inexpensive method.

J. Frhlich and H. Knig (1999) Rapid isolation of single microbial cells from mixed natural and laboratory populations with the aid of a micromanipulator. System. Appl. Microbiol. 22, 249-257 .


'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. High-Throughput Isolation of Total RNA
3. Simple Isolation of RNA from Tissue and Cultured Cells
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
6. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
7. Mouse Tail Genomic DNA Isolation Protocol(1)
8. Genomic DNA Isolation Protocol(1,2,3)
9. Basic Plasmid DNA Isolation Protocol
10. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
11. RNA-free Plasmid DNA Isolation Protocol
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/22/2017)... SPRING, Md. and RESEARCH TRIANGLE PARK, N.C., Feb. 22, ... UTHR ) today announced its financial results for ... "Our annual 2016 financial results reflect ... and earnings exceeded $700 million," said Martine Rothblatt, ... "These financial results strengthen our ability to develop ...
(Date:2/21/2017)... ... February 21, 2017 , ... During HIMSS ... digital health applications, announced a partnership with Redox, a leader in cloud-based healthcare ... many clinical systems while keeping data secure in the cloud. , The digital ...
(Date:2/21/2017)... ... February 21, 2017 , ... Cancer diagnostics workflow solution ... its executive team to lead the development and commercialization of its Cancer Information ... the diagnosis and treatment of cancer. The CIDT addresses the need for curated ...
(Date:2/21/2017)... ... February 21, 2017 , ... ... manufacturing facility at its headquarters laboratory in Poway, California. Based upon 12 ... of both in-house personnel and consultants, VetStem constructed and validated a state-of-the-art GMP ...
Breaking Biology Technology:
(Date:1/31/2017)... , Jan. 31, 2017  Spero Therapeutics, ... therapies for the treatment of bacterial infections, today ... of antibacterial candidates from Pro Bono Bio Ltd ... prevalence of multi-drug resistant forms of Gram-negative bacteria.  ... Anti Infectives Ltd, a PBB group company. ...
(Date:1/25/2017)... , Jan. 25, 2017 The Elements of ... (IAM) lifecycle is comprised of a comprehensive set ... purpose of maintaining digital identities and providing a ... applications. There are significant number of programs opted ... to time by optimizing processes and changing policies. ...
(Date:1/21/2017)... -- Research and Markets has announced the addition of ... their offering. ... The global voice recognition biometrics market to grow at a CAGR ... covers the present scenario and the growth prospects of the global ... the report considers the revenue generated from the sales of voice ...
Breaking Biology News(10 mins):