Prototheca stagnora

平台编号：bio-103873

 Prototheca stagnora Cooke 拉丁名

ATCC? 16528 编号

Strain Designations 菌株别名 SEC-L-1690 [UTEX 1443]

Biosafety Level 安全等级 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 分离基物  ludge, Lebanon, OH, 1961

Product Format 提供形式 frozen

Type Strain 模式菌株 yes

Medium 培养基 ATCC? Medium 28: Emmons' modification of Sabouraud's agar

Growth Conditions 生长条件 

Temperature 培养温度 : 25.0°C

Duration: axenic

Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface.

Subcultivation

Protocol: ATCCNO: 16522 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium to containing 12% sucrose to the freeze-dried inner shell vial. Once completely rehydrated, aseptically transfer the material to a 100 mm agar plate of the appropriate medium and evenly distribute the material over the surface of the agar with a spread bar. Subculture 4-6 weeks when incubated at 25C. Subculture 6-12 months when incubated at 18C To subculture, transfer a loopful of material to a fresh plate and spread evenly over the surface.

Cryopreservation

1.   Harvest cells from a culture which is at or near peak density by adding 3.0-5.0 ml fresh ATCC medium 28 broth to the slant or plate and washing cells into suspension.  It may be helpful to rub the surface of the agar with a spread bar or inoculating loop to detach adhering cells.

2.   Adjust the concentration of cells to 2 x 107/ml with fresh broth medium, then dilute to half this concentration by adding an equal amount of a 20% (v) sterile solution of either DMSO or glycerol in fresh ATCC medium 28 broth.

3.   Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).  The time from mixing of the cell preparation and the cryoprotective solution to the start of the cooling cycle should be no less than 15 min and no greater than 30 min.

4.   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.)   

5.   The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80 and -70°C for no longer than one week.

6.   To establish a culture from the frozen state place an ampule in a water bath set at 35°C until thawed (2-3 min). Immerse the ampule enough to cover only the frozen material.  Do not agitate the ampule.

7.   Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and add to

a fresh slant of ATCC medium 28 or the surface of an agar plate of ATCC medium 28.

8.   Maintain as described above.

Name of Depositor WB Cooke

Year of Origin 1961

References 参考文献 

Pore RS. Prototheca taxonomy. Mycopathologia 90: 129-139, 1985.

Cooke. J. Elisha Mitchell Sci. Soc. 84: 217-220, 1968.

type strain