Protostelium mycophaga Olive et Stoianovitch (ATCC® PRA-154)

Organism: Protostelium mycophaga Olive et Stoianovitch  /  Depositor: FW Spiegel

Permits and Restrictions

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Strain Designations Type
Application
Phylogenetic analysis
Biosafety Level 1
Isolation
plant (Phragmites australis (Cav.) Steudel (Phragmites communis, common reed) dead inflorescence, New Jersey meadowland)
New Jersey, United States
Isolation date: 1959
Type Strain no
Comments
Phylogenetic analysis
Medium ATCC® Medium 2432: wMY (weak Malt Yeast Extract)
Growth Conditions
Temperature: 15-20.0°C
Growth condition: Grown with Rhodotorula mucilaginosa ATCC MYA-3510 as food source
Cryopreservation
Cryoprotective Solution

DMSO                                                                                    1.5 ml

Fresh growth medium w/o bacteria                                 8.5 ml

1.     Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2.    Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryl's solution) and washing cells into suspension.  Rub the surface of the plate with a spread bar to detach adhering trophozoites.

3.     Adjust the concentration of cells to at least 2 x 104/ml in fresh medium.

4.     Mix the cell preparation and the cryoprotective solution in equal portions.

5.     Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6.     Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  

7.     Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8.     To establish a culture from the frozen state place the vial in a 35°C water bath.  Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.   Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 2432.  Distribute the material evenly over the plate using a spread bar.

9.     Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

10.   Follow the protocol for maintenance of culture.

Name of Depositor FW Spiegel
Special Collection eumycetozoan
Year of Origin 1959
References

Phragmites australis (Cav.) Steudel (Phragmites communis, common reed) dead inflorescence, New Jersey meadowland

Olive LS. The Protostelida - a new order of the Mycetoozoa. Mycologia 59: 1-29, 1967.

Olive LS, Stoianovitch C. Two New Members of the Acrasiales. Bull. Torrey Bot. Club 87: 1-20, 1960.

Shadwick LL, et al. Eumycetozoa=Amoebozoa?: SSUrDNA phylogeny of protosteloid slime molds and its significance for the amoebozoan supergroup. PLoS ONE 4(8): e6754, 2009.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation