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Hartmannella limax (Dujardin) Page
Hartmannella limax (Dujardin) Page
規(guī)格:
貨期:
編號:B227505
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 Hartmannella limax (Dujardin) Page
商品貨號 B227505
Strain Designations F-53
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.

Product Format dried
Type Strain no
Medium ATCC® Medium 997: Fresh water ameba medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Enterobacter aerogenes ATCC 13048 and with another hartmannellid
Protocol: ATCCNO: 30939 SPEC: Aseptically add 1 ml of sterile distilled water to the inner vial, remove the filter paper with a pair of forceps, and place it in the center of an agar plate of medium 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: 30939 SPEC: Aseptically add 1 ml of sterile distilled water to the inner vial, remove the filter paper with a pair of forceps, and place it in the center of an agar plate of medium 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
Cryoprotective Solution

DMSO ?????????????????????????????????????????????????????????????????????????????????? 1.5 ml

Dryl's solution (or similar)????????????????????? ??????????????????????????? 8.5 ml

1.???? Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2. ?? Harvest cells from a culture which is at or near peak density by adding 5 ml ATCC medium 5080 (Dryl's solution) and washing cells into suspension.? Rub the surface of the plate with a spread bar to detach adhering trophozoites.

3. ??? Adjust the concentration of cells to at least 2 x 106/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. 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.) ?

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

8.? ?? To establish a culture from the frozen state place the vial 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.?? Immediately after thawing, aseptically transfer the contents of the ampule to the center of a fresh plate of ATCC medium 997.? Distribute the material evenly over the plate using a spread bar.

9.     Wrap the entire edge of the plate with parafilm and incubate upright at 25°C.

Follow the protocol for maintenance of culture.

Name of Depositor JL Griffin
References

Page FC. Hartmannella limax: the original limax amoeba?. Trans. Am. Microsc. Soc. 88: 199-204, 1969. PubMed: 5798355

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