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| C O N T E N T S |
| C-Fern Home |
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| Introduction C-Fern Nutrient Medium Culture Conditions Gametophyte Culture Gametophyte Observations Sporophyte Culture Spore Mutagenesis Bibliography |
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Light Stand Growth Pod DNA Extraction C-Fern T-shirt Who's Who Selection and mutants Media formulation error Image gallery |
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National Science Standards Gametophyte development Student research questions Research in the classroom Research with C-Fern Brief description of C-Fern C-Fern manipulation Root tropism? |
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| Investigations Mutant & wild type spores Manual & Culture Instructions Supplies - Domes, media |
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of C-Fern is supported by the National Science Foundation (NSF-DUE) |
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Copyright
© 1997-2000
Thomas R. Warne and Leslie G. Hickok. All rights reserved. |
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C-Fern
Web Manual - Spore Inoculation
Spore Sowing and Density Calculations
Through painstaking analyses, we have determined that there are about 1250 wild type C-Fern spores per mg. One mg of C-Fern spores will inoculate 3.5 small Petri dishes with about 0.28 mg or 300+ spores per dish. For a typical 60x15 mm (about 22.5 cm2) Petri dish, this inoculation rate is about 15 spores per cm2 of agar surface. At substantially below this density, sex expression stabilizes and gametophyte growth is relatively unaffected. If density is important, the actual area of dishes should be empirically determined since nominal dimensions are substantially different for most dishes. Since known quantities of water are used to suspend spores for sowing and this suspension is delivered by drops from a Pasteur pipet, it is important to determine the drop volume from your given brand of glass or plastic Pasteur pipet. We use a plastic transfer pipet which, for a 1 ml inoculation volume will sow about 8 dishes at 3 drops per dish. If using other pipets, drop volume must be measured and factored in to achieve controlled sowings. For liquid culture, spores can be sown into media without agar at a rate of 3 to 10 mg per 100 mL. Cultures should be placed on shaker at 50-100 rpm, depending on the volume of solution. If not shaken, cultures will rapidly fertilize when gametophytes become sexually mature. |
