For classroom or independent investigations, the following materials and supplies may be necessary. Culture Media and Media Preparation (per liter of prepared medium) inorganic salts for Basic C-Fern Nutrient Media OR pre-packaged OR prepared Basic C-Fern Nutrient Medium If using inorganic salts, supplies for weighing and preparing stock solutions (see Basic C-Fern Nutrient Medium recipe) […]
An appropriate culture environment helps regulate temperature, humidity, evaporation, and helps give consistent results. Culture Containers Once cultures are inoculated with spores, place them into an appropriate culture container. Culture Dome – consisting of clean plastic greenhouse trays covered with transparent humidity domes. Culture Domes should be thoroughly clean to prevent contamination of cultures by […]
Simple lighting setups can provide adequate illumination for gametophyte growth and can help control culture temperature Figure 1. C-Fern Growth Pod illuminated by a 15-watt screw-in fluorescent bulb placed in a small (ca. 6 in) reflective fixture. Once cultures have been inoculated with spores, place them into an appropriate environment that will provide a constant temperature, 28 […]
Preparation of C-Fern Basic Nutrient Solution using Powdered Nutrient Medium. Materials Powdered Basic C-Fem Medium 1 Liter volumetric flask Distilled or deionized water 2 Liter Erlenmeyer Flask{s) (1 flask per liter of medium) Agar (BactoAgar®) Magnetic Stir Plate and Magnetic Stir Bar pH Meter (for pH adjustment) 0.1 M NaOH Solution Foil (to cover flask […]
Preparation of C-Fern Basic Nutrient Medium – 3 Stock Solution Method. This method has been modified from the traditional 6-Stock Solution method to make final nutrient solution preparation easier. Preparation of Stock Solutions Materials Macro-, micronutrient and Fe salts (Table 1) Distilled or deionized water 1 L volumetric flask(s) Microbalance Magnetic stir plate and stir […]
Preparation of Traditional C-Fern Basic Nutrient Medium – 6 Stock Solution Method
Technique for pouring of agar-solidified nutrient medium into Petri dishes.
Using a Microwave to Prepare Basic C-Fern Powdered Medium Makes C-Fern Even Easier to Use!C-Fern users who do not have access to an autoclave can successfully use a microwave to prepare Basic C-Fern Powdered Medium. Although the lower temperatures and shorter times of microwaving cannot guarantee sterility to the degree that autoclaving does, this method […]
Establish sterile cultures with a simple method for the surface sterilization of C-Fern spores. All spore sterilization and sowing procedures should be performed on a sterile bench or laminar flow hood. Figure 2. We recommend that C-Fern spores be surface sterilized in 6 ml plastic tubes with conical bottom and screw caps and handled with […]
Inoculation or “Sowing” of C-Fern Spores on the agar surface. Figure 1. Supplies needed for sowing C-Fern spores Materials Surface sterilized C-Fern spores in a graduated spore vial. (Wild type (RNWT1) and mutant spores are available as pre-sterilized class sized and bulk unsterilized units. Bulk unsterilized spores and spores collected directly from sporophytes should be […]
Spore Sowing and Density Calculations Through painstaking empirical analyses, we determined that there are about 1250 wild type C-Fern spores per mg, i.e. each spore weighs about 0.8 µg. One mg of C-Fern spores will inoculate 3.5 small Petri dishes with about 0.24 mg or 300+ spores per dish. For a typical 60 x 15 mm (about 22.5 cm²) Petri […]
C-Fern