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Paramecium tetraurelia Sonneborn 拉丁名

(ATCC? 30763?) 統(tǒng)一編號

Strain Designations 菌種別名 stock d4-89 

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 mutant cross, 1970 

Product Format 提供形式 test tube 

Type Strain 模式菌株 no 

Genotype cl2 mtB(o) nd3a sclc2 ts401

Medium 培養(yǎng)基 ATCC? Medium 802: Sonneborn's Paramecium medium 

Growth Conditions 生長條件 

Max Temperature 最高溫度 : 27.0°C 

Min Temperature 最低溫度 : 13.0°C 

Protocol: ATCCNO: 30300 SPEC: This strain is shipped as a growing test tube culture. Upon arrival, remove test tube from sealed plastic envelope, remove plastic seal from cap, and loosen the cap one half turn. 

Add 1.0 ml of ATCC medium 802 bacterized with Enterobacter aerogenes ATCC 13048 twice weekly. 

When the tube is filled to within one inch of the top, decant leaving 5.0 ml in the original tube. 

Subcultures are established by transferring 0.5 ml of a growing culture to 5.0 ml of bacterized ATCC medium 802 in a 20 x 120 mm test tube. 

Subcultivation Protocol: ATCCNO: 30300 SPEC: This strain is shipped as a growing test tube culture. Upon arrival, remove test tube from sealed plastic envelope, remove plastic seal from cap, and loosen the cap one half turn. 

Add 1.0 ml of ATCC medium 802 bacterized with Enterobacter aerogenes ATCC 13048 twice weekly. When the tube is filled to within one inch of the top, decant leaving 5.0 ml in the original tube. 

Subcultures are established by transferring 0.5 ml of a growing culture to 5.0 ml of bacterized ATCC medium 802 in a 20 x 120 mm test tube. 

Cryopreservation Cryoprotective Solution DMSO 1.5 ml Fresh growth medium w/o bacteria  7.5 ml MgCl2 (0.5 mM) 0.5 ml CaCl2 (0.5 mM) 0.5 ml 

1.Mix the components in the order listed.  Before adding the MgCl2 and the CaCl2 allow the solution to return to room temperature.  When the medium is added to the DMSO the solution will warm up due to chemical heat. 

2.Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 200 x g for 1 min. 

3.Adjust the concentration of cells to 2 x 105/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. 

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

8.To establish a culture from the frozen state add 1.0 ml ATCC medium 802 to the frozen ampule and place it 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. 

9.Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate onto the surface of an ATCC medium 919 (non-nutrient agar) plate containing an overlay of 15.0 ml of bacterized ATCC medium 802. 

10.Incubate at 25°C. 

11.Once the culture is established, transfer 0.5 ml to 5.0 ml of bacterized ATCC medium 802. 12. Follow the protocol for maintenance of culture. 

Name of Depositor 寄存人 TM Sonneborn 

Chain of Custody 來源國家 ATCC <<--TM Sonneborn<<--J. Dilts 

Year of Origin 1970 

References 參考文獻 Handbook of genetics. vol. 2New York: Plenum Press; 1975.