Snow Algae

From Protists

Background

These cultures were originally from Ron Hoham and are currently in the UTEX culture collection. We were storing them and maintaining them for use in a collaboration between Ron Hoham (now retired from Colgate) and Dr. Jim Leebens-Mack (plant systematist at UGA). However, most were contaminated with fungi and/or lost because they were killed when the walk-in broke over the summer of 2007 and/or they were not transferred often enough and the agar dried out.

Snow Algae Media Recipe

1. Measure 500mL DI H₂O in a LIVE graduated cylinder
2. Add appropriate amount of all chemical stock solutions (Table 1) to the graduated cylinder
3. Mix until all chemicals are dissolved in the solution (should be yellow-golden color)
4. Bring volume in graduated cylinder up to 1000mL with DI H₂O
5. Check pH with pH meter and adjust solution to pH ~5.1-5.3 (using HCl or NaOH as needed)
6. Dispense 500mL into two 1L LIVE media bottles and add 8g of LB Agar to each bottle
7. Autoclave media bottles on the liquid cycle with at least 30 minutes of sterilization time
8. Pour contents of media bottles into bottom half of petri dishes (~3/4 full) when media is cool enough (but not hardening!)
9. Store plates upside-down petri dish bags (taped closed and labeled as Snow Algae Plates) in the walk in cooler (~5C)

Table 1. Chemical stock solutions to be added to snow algae and how to make the stock solutions.

Chemical Compound	Stock Solution Composition	Volume of Stock Solution to Add (mL)
CaCl2	0.5g per 100mL	1
NaNO3	2.5g per 100mL	1
NH4Cl	0.5g per 100mL	1
CaSO4 x 2H2O	0.4g per 100mL	1
MgSO4 x 7H2O	0.5g per 100mL	1
NaSiO3 x 9H2O	0.2g per 100mL	1
Fe (as EDTA)	see notes below	250
Trace Elements	see below (Table 2 and notes)	10
K2HPO4	3.48g per 100mL (0.2M)	2
KH2PO4	2.72g per 100mL	98

Snow Algae Agar Plates

• measure out 525mL Snow Algae Media (above) into a LIVE graduated cylinder
• add 10.5g of Agar (not agarose!!) to make a 2% weight/volume agar concentration
• distribute the mixture into a LIVE media bottle(s)
• heat the mixture on the LIVE hot/stir plate to dissolve the agar completely
• Autoclave the mixture in the bottles on the liquid cycle for the appropriate time