| 产品名称： |
Mastigamoeba aflagellifera |
| 商品货号： |
TS147222 |
| Strain Designations： |
AF065-Y |
| 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： |
Soil of a rice field, Kanagi, Goshogawara city, Aomori pref., Japan, Dec. 1, 2006 |
| Product Format： |
frozen |
| Storage Conditions： |
Frozen Cultures:
-70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures:
2-8°C
Live Cultures:
See Protocols section for handling information |
| Type Strain： |
yes |
| Medium： |
ATCC® Medium 2832: Reduced YPD Medium
ATCC® Medium 2154: LYI Entamoeba medium
|
| Growth Conditions： |
Temperature: 10°C to 30°C
Atmosphere: Microaerophilic
Culture System: Axenic |
| Cryopreservation： |
Reagents
Cryoprotective Solution
DMSO, 2.0 mL
Fresh growth medium, 8.0 mL
Harvest and Preservation
-
Harvest cells from several cultures that are in the late logarithmic to early stationary phase of growth.xa0 Place culture vessels on ice for 20-30 min.
-
Vigorously invert tubes 20-30 times or as necessary to sufficiently detach cells, then centrifuge at 500 x g for 5 min.xa0 Handle cultures promptly after centrifugation to avoid the amoebae reattaching to the culture tubes.xa0 Note:xa0 Increased yield may be obtained by using a sterile cotton swab to rub the inside surface of culture tubes both before and after centrifugation.xa0 Use of a refrigerated centrifuge will aid in preventing reattachment of cells to culture tubes during or immediately following centrifugation.xa0xa0xa0xa0xa0xa0xa0xa0
-
Adjust the concentration of cells to between 5 x 105/mL - 5 x 106/mL using reduced medium (i.e., supernatant).
-
Mix the cell preparation and the cryoprotective solution in equal portions. xa0Invert the tube several times to obtain a uniform cell density.
-
Dispense 0.5 mL aliquots into 1.0 - 2.0 ml plastic sterile cryules (special plastic vials for cryopreservation).
-
Place the vials in a controlled rate freezing unit.xa0 Use the following cooling cycle: From room temperature cool at -10°C/min to the heat of fusion; from the heat of fusion toxa0-40°C, cool at -1°C/min.xa0xa0At -40°C plunge into liquid nitrogen.xa0 The cooling cycle should be initiated no less than 15 and no more than 30 minutes after the addition of DMSO to the cell preparation.
-
Store ampules in a liquid nitrogen refrigerator until needed.
-
To establish a culture from the frozen state, place an ampule in a 35°C water bath, until thawed (2-3 min).xa0 Immerse the vial just sufficiently to cover the frozen material.xa0 Do not agitate the ampule.
-
Aseptically transfer contents of thawed ampule to a glass, rubber-seal screw-capped tube containing 14-15 mL ATCC Medium 2832.
-
Screw cap on tightly and incubate on a 15° horizontal slant at 10-30°C (20-25°C recommended for routine cultivation).xa0 Observe the culture daily and transfer when many trophozoites are observed.
|
| Name of Depositor： |
A Tonouchi |
| Chain of Custody： |
ATCC <-- A Tonouchi |
| References： |
Uchimura Y, et al. Mastigamoeba aflagellifera sp. nov. isolated from the soil of a rice field in Japan. (in preparation for submission)
|
| Cross References： |
Nucleotide (GenBank) :
AB55091
ssu rRNA gene sequence of strain AF065-Y
|
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