Effectively mutagenize C-Fern spores for use in selection protocols. WARNING EMS IS A POTENT MUTAGEN Use proper clothing, gloves, containment trays, supplies, and equipment. Sodium thiosulfate solution should be used to neutralize any EMS spills and to neutralize treatment solutions after mutagenesis. All mutagenesis procedures should be carried out in an appropriate exhaust/fume hood. Materials […]
Have an old x-ray machine lying around. Use x-rays to effectively mutagenize spores for selection protocols Materials X-ray machine Petri Dish, glass Plastic sandwich bag C-Fern spores Materials and supplies for spore sterilization and culture inoculation Methods SOAK SPORE Soak spores for 24 hours prior to sterilization and irradiation. STERILIZE SPORES Sterilize using standard methods […]
Create semi-permanent microscope slides of gametophytes using a mixture of a stain in a combination clearing – fixative – preservative agent. Semi-permanent slides suitable for later observations or archival documentation can be prepared by mounting gametophytes in a Hoyer’s/Acetocarmine mixture. Hoyer’s Medium clears and preserves tissue; acetocarmine is one of the few stains stable in […]
Designing an appropriate selection procedure is important for successfully identifying mutants. The identification of appropriate selection conditions typically requires careful dose-response curves to determine the correct concentration of a toxin (for example, selections using herbicides, FuDR, etc.) that results in lethality or sufficient and distinctive inhibition of gametophyte growth or development. Several factors may need […]
A Method for Observing Ceratopteris richardii SpermHeather Cousino and Jun Tsuji, Siena Heights University, Department of Biology, 1247 E. Siena Heights Drive, Adrian, Michigan 49221-1796, USA ABSTRACT The fern Ceratopteris richardii produces motile sperm that are challenging to observe and study due to their rapid speed. To assess the effectiveness of Protoslo in restricting the […]
DNA Extraction Procedure for Ceratopteris – DNA Preparation AttemptsTim Chipman (July 1995) I have attempted a number of total nucleic acid preps using C. richardii prothalli and have had some degree of success. It appears that the limiting factor in this protocol is the amount of tissue readily available. I have been culturing 2.5L batches […]
DNA Extraction Procedure for Ceratopteris – Modified QIAGEN Genomic-tip ProcedureLeslie G. Hickok 4/96 The following procedure results in yields of about 25 ug per g fresh weight of 2 week old gametophytic tissue. Grind 2-3 g gametophytes in liquid nitrogen with a mortar and pestle. Harvest tissue from ca. 2 week old liquid cultures (50 […]
A Method of Producing STS (Sequence Tagged Site) markers from RAPDs (Random Amplified Polymorphic DNAs) in Ceratopteris richardii Cora Chaffin (Provine High School, Jackson, Mississippi) and Robert Hamilton (Mississippi College, Clinton, Mississippi) INTRODUCTION Genes are arranged in specific sequences along chromosomes, with each gene occupying its own particular position (locus) on a chromosome. Together the […]
A rapidly developing C-Fern strain can improve studies of plant life cycles and development. C-Fern Express is a select strain of an uncommon fern, Ceratopteris. It was derived by Dr. Les Hickok from two Japanese varieties of C. thalictroides that exhibited very rapid sporophyte development. Its novel features and very rapid development time make it […]