Salpingoeca sp. (ATCC® 50931)

Strain Designations: BSE-01190019  /  Depositor: TA Nerad  /  Biosafety Level: 1

Permits and Restrictions

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Strain Designations BSE-01190019
Biosafety Level 1
Isolation
dry salt marsh sediment, Hog Island, eastern shore of Virginia, 2000
Product Format frozen
Type Strain no
Medium Medium 1525: Seawater 802 medium
Growth Conditions
Temperature: 25.0°C
Duration: grown with Klebsiella pneumoniae subsp. pneumoniae ATCC 700831 and mixed bacteria
Protocol: To subcultivate, virgorously agitate a T-25 flask at or near peak density and aseptically transfer a 0.5-ml aliquot to a fresh T-25 flask containing 10 ml of bacterized ATCC medium 1525.
Subcultivation
Protocol: To subcultivate, virgorously agitate a T-25 flask at or near peak density and aseptically transfer a 0.5-ml aliquot to a fresh T-25 flask containing 10 ml of bacterized ATCC medium 1525.
Cryopreservation
Cryoprotective Solution

DMSO                                                                                    2.0 ml

Fresh growth medium w/o bacteria                                 8.0 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 that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.

3.     Adjust the concentration of cells 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, do not leave in water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 ml ATCC medium 1525 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC®  700831).

9.       Incubate at 25°C with the cap screwed on tightly.

10.     Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 ml to 10.0 ml of bacterized ATCC medium 1525.

11.     Follow the protocol for maintenance of culture.

Name of Depositor TA Nerad
Year of Origin 2000
References

Thomas A Nerad, personal communication

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation