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Crypthecodinium cohnii (Seligo) Javornicky 菌株拉丁名

 (ATCC? 30556?) 菌株編號

Strain Designations 菌株別名 : To (Tolo) /

Depositor: CA Beam, M Himes /

Biosafety Level 生物安全等級 : 1

Strain Designations 菌株別名 To (Tolo)

Application 用途 Biofuel production

Biosafety Level 生物安全等級 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 分離源 Shoreline organic debris, Tolo Harbor, Hong Kong, 1976

Product Format 提供形式 frozen

Storage Conditions 保藏方法

Frozen Cultures 冷凍培養(yǎng) : -70℃ for 1 week; liquid N2 vapor for long term

storage 保藏規(guī)格 Freeze-dried

Cultures: 2-8℃

Live Cultures 活菌 : See Protocols section for handling information

Type Strain 模式菌株 no

Comments 注釋 Minor sibling species Genetic diversification Sexuality and meiosis

Medium 培養(yǎng)基 ATCC? Medium 460: A2E6 medium

Growth Conditions 生長條件

Temperature 培養(yǎng)溫度 : 20℃ to 25℃

Culture System: Axenic Cryopreservation Harvest and Preservation

1.Harvest cells from cultures which are at or near peak density.Aseptically transfer cells to 15 mL plastic centrifuge tubes and centrifuge at ~150 x g for 5 min.

2.Adjust the concentration of cells to 2 x 106/mL with fresh medium, then dilute to half this concentration by adding an equal amount of a 15% (v/v) sterile glycerol solution in fresh medium.

3.Dispense in 0.5 mL aliquots into 1.0 mL to 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation). The time from mixing of the cell suspension and the glycerol solution, before the cooling cycle begins, 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℃/min to -40℃. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40℃ plunge ampules into liquid nitrogen.

Alternatively, place the vials in a Nalgene 1℃ freezing apparatus. Place the apparatus at -80℃ for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1℃/min.)

5.The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130℃ are stabile indefinitely. Those stored at temperatures above -130℃ are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80℃ and -70℃ 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℃. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.

7.Immediately after thawing, do not leave in the water bath; aseptically remove the contents of the ampule and transfer to a fresh tube (or T25 flask) containing 5 mL of ATCC Medium 460. Incubate culture (if in a tube, incubate vertically) at 20℃ to 25℃ with the cap loosened one-half turn. Subculture every 10 to 14 days.

Name of Depositor 寄存人 CA Beam, M Himes

Year of Origin 1976

References 參考文獻 Beam CA, Himes M. Sexuality and meiosis in dinoflagellates. In: Beam CA, Himes M, Biochemistry and physiology of protozoa, 2nd ed.3 New York Academic Press: 171-206, 1980.

Beam CA, Himes M. Sexual isolation and genetic diversification among some strains of Crypthecodinium cohnii-like dinoflagellates evidence of speciation. J. Protozool. 24: 532-539, 1977.

Iizuka T, et al. Marine micro-algae food material containing docosahexaenoic acid, food containing the same and manufacturing method therefor. US Patent 5,547,699 dated Aug 20 1996

Japanese Patent 7,008,221 Salt GW. Changes in the cell volume of Didinium nasutum during population increase. J. Protozool. 22: 112-115, 1975.

Seafood products having good taste containing marine microalgae, particularly Crythecodinium cohnii containing docosahexaenoic acid. Japanese Patent 7,008,221