Naegleria minor (Dobson et al.) De Jonckheere and Brown
商品貨號
B179902
Deposited As
Willaertia minor Dobson et al.
Strain Designations
SWL WT-043
Biosafety Level
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Isolation
freshwater sediment, Berry Springs, Northern Territory, Australia, 1990
Product Format
dried
Type Strain
no
Medium
ATCC® Medium 997: Fresh water ameba medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Escherichia coli ATCC 11775
Protocol: ATCCNO: 30133 SPEC: This strain is shipped as a dried preparation. Aseptically add 1 ml of sterile distilled water to the inner vial, remove the filter paper pellet with a pair of forceps, and place it in the center of an agar plate of ATCC medium 711 or 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days.
Subcultivation
Protocol: ATCCNO: 30133 SPEC: This strain is shipped as a dried preparation. Aseptically add 1 ml of sterile distilled water to the inner vial, remove the filter paper pellet with a pair of forceps, and place it in the center of an agar plate of ATCC medium 711 or 997. Add the liquid remaining in the vial to the plate and spread it evenly over the surface of the plate. Incubate the plate at 25C. Trophozoites (amebae) should be evident within 2-3 days.
Cryopreservation
1. ? Allow the cells to encyst.? To detach cysts from the plate flush the surface with 5 ml fresh ATCC medium 1323 (Page's Balanced Salt Solution).? Rub the surface of the plate with a spread bar to detach adhering amoebae.
2.?? Transfer the cyst suspension to a sterile centrifuge tube.
3. ? If the cyst concentration does not exceed 2 x 106 cysts/ml adjust the suspension to that concentration.? To adjust the concentration, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield 2 x 106.
4.?? While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows:? Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath.? Allow the DMSO to solidify.? Add the required volume of refrigerated medium.? Dissolve the DMSO by inverting the tube several times.?
Name of Depositor
BS Robinson
Year of Origin
1990
References
De Jonckheere JF. A century of research on the amoeboflagellate genus Naegleria. Acta Protozool. 41: 309-342, 2002.
