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Cyanidioschyzon merolae P. De Luca, R. Taddei & L. Varano

PRA-402

Product category
Protists
Product type
Algae
Classification
Cyanidiophyceae, Cyanidiales, Cyanidiaceae
Strain designation
10D
Genome sequenced strain
Yes
Isolation source
Hot spring
Geographical isolation
Italy; Sardinia
Product format
Frozen
Storage conditions
-80°C or colder for 1 week, vapor phase of liquid nitrogen for long-term storage
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information

Characteristics

Extremophile type
Acidophile
Thermophile
Comments
Genome sequencing strain

Handling information

Medium
Instruction for complete medium
Media: ATCC Medium 2861: Acidic Allen's medium

Alternate Media: ATCC Medium 616: Medium BG-11 for blue-green algae with pH adjusted to 2.3 with H2SO4.
Temperature
42°C
Handling procedure
Storage and Culture Initiation
Frozen ampules packed in dry ice should either be thawed immediately or stored in liquid nitrogen.  If liquid nitrogen storage facilities are not available, frozen ampoules may be stored at or below -70°C for approximately one week.  Do not under any circumstance store frozen ampules at refrigerator freezer temperatures (generally -20°C).  Storage of frozen material at this temperature will result in the death of the culture.
  1. To thaw a frozen ampule, place it in a 35°C water bath until thawed (2-3 min).  Immerse the ampule enough to cover only the frozen material.  Do not agitate the ampule.
  2. Immediately after thawing, aseptically transfer contents to a 16 x 125 mm screw-capped test tube containing 5.0 mL of ATCC medium 2861. Incubate the tube on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 42°C under constant light.
  3. Examine the culture using an inverted microscope for the presence of green microalgae. Growth of the strain may take several weeks to establish.
Culture maintenance
  1. Screw the cap on tightly and vigorously agitate the culture.
  2. Aseptically transfer a 0.5 mL aliquot to 5 mL of fresh medium in a 16 x 125 mm screw-capped test tube.
  3. Screw caps on loosely (loosened one-half turn) and incubate on a 15° horizontal slant at 42°C under constant light.
  4. Subculture every 2-3 weeks.
Cryopreservation
  1. Harvest cells from a culture which is at or near peak density by centrifugation at ~800 x g for 10 min.
  2. Adjust concentration of cells to 2 x 107 - 2 x 108 mL in fresh growth medium.  If the concentration is too low, centrifuge at ~800 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired cell concentration.
  3. While cells are centrifuging, prepare a 10% (v/v) solution of sterile methanol in fresh growth medium. 
  4. Mix the cell suspension and the methanol solution in equal portions. The final concentration will be 107 – 108 cells/mL and 5% (v/v) of methanol. The time from the mixing of the cell preparation and methanol stock solution before the freezing process is begun should be no less than 5 min and no more than 15 min.
  5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.
  6. Place the vials in a controlled rate freezing unit.  From room temperature cool at -1°C/min to -40°C.  If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion.  At -40°C plunge 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. Store in either the vapor or liquid phase of a nitrogen refrigerator.
  8. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after it is thawed.
  9. Remove the vial from the water bath immediately after thawing.  Aseptically transfer the contents of the ampule into 5 mL of fresh ATCC medium 2861. 
  10. Incubate the tube on a 15° horizontal slant with the cap screwed on loosely (loosened one half turn) at 42°C under constant light.
  11. Maintain as described above.  

History

Deposited as
Cyanidioschyzon merolae
Depositors
T. Kuroiwa
Chain of custody
ATCC <-- T. Kuroiwa
Type of isolate
Environmental

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Ohta N, et al. Structure and organization of the mitochondrial genome of the unicellular red alga Cyanidioschyzon merolae deduced from the complete nucleotide sequence. Nucleic Acids Res. 26(22): 5190-5198, 1998. PubMed: 9801318

Ohta N, et al. Complete sequence and analysis of the plastid genome of the unicellular red alga Cyanidioschyzon merolae. DNA Res. 10(2): 67-77, 2003. PubMed: 12755171

Matsuzaki M, et al. Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D. Nature 428(6983): 653-657, 2004. PubMed: 15071595

Nozaki H, et al. A 100%-complete sequence reveals unusually simple genomic features in the hot-spring red alga Cyanidioschyzon merolae. BMC Biol. 5: 28, 2007. PubMed: 17623057

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