Organisms for AToL Collaboration
From Protists
Last updated on 20Feb09 by CC
- To maintain cultures, use 25cm small flask and transfer once per week, with a total volume of 15 mL per small flask
- To grow and harvest cultures, use 150cm large flasks (except Diplonemids, use many small flasks) and transfer as noted in detail in the flask culture columns, with a total volume of 125 mL per large flask with exceptions.
- Can use sterile cell scrapers for all cultures to harvest cells except Diplonemids and Peranema(just agitate gently before pouring out to spin down)
- We use sterile Corning tissue culture, canted neck, plug seal flasks (close all the way) because they are cheaper and cut down on contamination, but vented flasks can be used for any of these cultures (Corning is apparently the best brand because it has the least leakage into the culture media)
- Some of these cultures used to be inoculated with either Enterobacter aerogenes (Ear; ATCC 13048) and/or Klebsiella pneumoniae (Kpn; ATCC 10273), but they aren't anymore although traces of those bacteria might remain in the cultures
- Some of these cultures have other organisms in the media to be noted (Hay, Cerophyll, Rice, Wheat, Barley, Soil, Egg Yolk, Heat-Inactivated Horse Serum, etc.)
- All flasks should be sterilely scrapped before spin downs or refreshing the media unless noted
- Be sure to wear gloves when working with the Diplonemids due to the Chloramphenicol and HI-HS. Also, wear gloves with the Peranema culture because the egg yolk and the cultures are axenic and MUST stay that way.
| 3-Letter Code | Organism Name | Culture Collection ID | Euglenozoan Clade | Media Used | Growth Temp (°C) | Small Flask Cultures | Large Flask Cultures | Bacterial Food History/Organisms from Media |
|---|---|---|---|---|---|---|---|---|
| BFP | Bodo from Peranema | Bodonids | Hay Infusion | 20 | refresh once a week | Hay | ||
| BST | Bodo saltans | CCAP 1907-2 | Bodonids | CCAP S/W | 15 | (spin down ~3-4 days) | (spin down ~3-4 days) | Soil |
| DMT | Dimastigella trypaniformis | ATCC 50263 | Bodonids | ATCC 802 | 25 | Start a new flask every 1-2 weeks; ~1mL from original culture + media to a total volume of 15 mL (spin down every ~2-3days) | Start a new flask every 1-2 weeks; ~10mL from original culture + media to a total volume of 125 mL (spin down every ~2-3days) | Kpn & Ear/Cerophyll |
| DI2 | Diplonema sp.2 | ATCC 50224 | Diplonemids | Artificial Seawater w/ 1% HI-HS | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add 1.5 mL HI-HS to flask with media [15 mL total volume] just prior to inoculating, swirl small flask, add ~5-10 drops of culture using a wide pipet tip (spin down every ~24-48hrs) | n/a | HI-HS |
| DIA | Diplonema ambulator | ATCC 50223 | Diplonemids | Artificial Seawater w/ 10% HI-HS & 1 mL Chloramphenicol | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add 1.5 mL HI-HS and 1 mL Chloramphenicol to flask with media [15 mL total volume] just prior to inoculating, swirl small flask, add ~510 drops of culture using a wide pipet tip (spin down every ~24-48hrs) | n/a | HI-HS |
| DIP | Diplonema papillatum | ATCC 50162 | Diplonemids | Artificial Seawater w/ 1% HI-HS & 1 mL Chloramphenicol | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add 150 uL HI-HS and 1 mL Chloramphenicol to flask with media [15 mL total volume] just prior to inoculating, swirl small flask, add ~5-10 drops of culture using a wide pipet tip (spin down every ~24-48hrs) | n/a | HI-HS |
| ENTO | Entosiphon sp. | contaminant in PERsc original culture | Euglenids | Hay Infusion w/ Barley | 15 | Add 3-4 grains of autoclaved barley to media; transfer to new small flask w/o transferring barley and add new barley after spin downs when barley has become dissociated (spin down ~4-7 days; refresh media every 3 weeks) | Add 10-15 grains of autoclaved barley to media; transfer to new large flask w/o transferring barley and add new barley after spin downs when barely has become dissociated (spin down ~4-7 days) | Hay, Barley |
| NBD | Neobodo designis (formerly Bodo designis) | CCAP 1951/1 | Bodonids | PE w/ Wheat | 25 | Add 3-4 grains of autoclaved wheat to media; transfer to new small flask w/o transferring wheat and add new wheat after spin downs when wheat has become dissociated (spin down ~3-4 days) | Add 10-15 grains of autoclaved wheat to media; transfer to new large flask w/o transferring wheat and add new wheat after spin downs when wheat has become dissociated (spin down ~3-4 days) | Wheat |
| PCT | Petalomonas cantuscygni | CCAP 1259/1 | Euglenids | ESNW w/ rice | 20 | ~1mL from original culture + media [total volume of 15 mL] + 4-5 autoclaved rice grains (spin down every ~2-3days); Make new flask each time. | ~10mL from original culture + media [total volume of 50 mL] + 20 autoclaved rice grains (spin down every ~2-3days); Make new flask each time. | Rice |
| PERsc | Peranema sp. (single cell) | Carolina Biological 131838 (not uniprotistan culture) | Euglenids | Hay Infusion or Knops w/ sterile Egg Yolk | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add media, add 1 10-100 uL tip (3-4 mm up the tip w/ sterile egg yolk) of sterile egg yolk, shake flask to break up egg yolk, add ~5 drops of culture using a wide pipet tip (~spin down 4-6 days; refresh weekly if you are not spinning down) | Start a new flask every time; Rock large flask ~5-6 times gently before transfering; Add media ~45 mL of media, add 3 10-100 uL tips (3-4 mm up the tip w/ sterile egg yolk) of sterile egg yolk, shake flask to break up egg yolk, add ~5 mL of culture (~spin down 4-6 days; refresh weekly if you are not spinning down) | Egg Yolk, Hay |
| PER | Peranema tricophorum | CCAP 1260/1b (from G. Burger) | Euglenids | Hay Infusion or Knops w/ sterile Egg Yolk | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add 15 mL media, add 1 10-100 uL tip (3-4 mm up the tip w/ sterile egg yolk) of sterile egg yolk, shake flask to break up egg yolk, add ~5-10 drops of culture using a wide pipet tip (~spin down 4-6 days) | Start a new flask every time; Rock large flask ~5-6 times gently before transfering; Add media ~45 mL of media, add 3 10-100 uL tips (3-4 mm up the tip w/ sterile egg yolk) of sterile egg yolk, shake flask to break up egg yolk, add ~5mL of culture(~spin down 4-6 days) | Egg Yolk, Hay |
| PLC | Ploeotia costatum | CCAP 1265/1 | Euglenids | CCAP ASWP w/ rice | 20 | Add 4-5 grains of autoclaved rice (spin down every ~1week-10days); Add 1 grain of rice every time you refresh (refresh every 3 weeks) | Add 10-20 grains of autoclaved rice (spin down every ~1week-10days); Add 3 grains of rice every time you refresh (refresh every 3 weeks) | Rice |
| PLO | Ploeotia oblonga | ATCC 50573 | Euglenids | optimizing Hay Infusion/ATCC 802 + Ear, +Kpn or +Ear&Kpn | 25 | (spin down every ~4days) | (spin down every ~4days) | Kpn, Hay |
| RBS | Rhynchobodo sp. | ATCC 50359 | Bodonids | CCAP S/W | 25 | (spin down every ~4days) | (spin down every ~4days) | Soil |
| RMN | Rhynchomonas nasuta AZ-4 | ATCC 50292 | Bodonids | ATCC 802 | 25 | Start a new flask every 1-2 weeks;(spin down ~3-4 days, refresh media once a week) | Start a new flask every 1-2 weeks;(spin down ~3-4 days, refresh media once a week) | Kpn & Ear/Cerophyll |
| RP1 | Rhynchopus euleeides (formerly Rhynchopus sp.1) | ATCC 50226 (from G. Burger) | Diplonemids | RPS_Ult + 10% HI-HS | 25 | Start a new flask every time; Rock small flask ~5-6 times gently before transfering; Add 1.5 mL of HI-HS + media [total volume of 15 mL] just prior to inoculating, swirl small flask, add ~5-10 drops of culture using a wide pipet tip (spin down every ~24-48hrs) | n/a | HI-HS |
