Category Archives: Protocols
Cultures preparartion for RNA extraction
1. Cut some mycelial plugs (like 5) from a 10 -14 days old PDA culture. 2. Inoculate with the agar plugs a flask with 100 ml of Fries medium 48 hours with shaking at 30 C. I use a … Continue reading
Recovery of C. graminicola from Root Infected Plants
Once a set of root inoculated plants reaches three weeks of age, transfer the plants to a 0.5L pot and fill using the green house soil. (The green house soil is a mixture of a of 1:2 store soil and … Continue reading
Root Inoculation Assay
Grow C. graminicola in Potato Dextrose Agar media for 2-3 weeks. Sterilize the surface of the seeds by following the Corn Seed Sterilization protocol. Wash vermiculite by placing it in a container for 2½ hours then pour out the water … Continue reading
Seed sterilization (of corn seeds for root inoculations)
1. Apply fungicide. Flint 50WG (Trifloxysfrobin) 0.8g fungicide/ Kg of seed 0.008g fungicide/ 10g seed in 0.5mL ddH2O 2. Allow fungicide to dry over night. 3. Rinse seeds with water. 4. Surface sterilize seeds by washing in a 70% ethanol … Continue reading
Genomic DNA Extraction From Fungi (maxi_DNA extraction)
Time: about 5-6 hours, Yield- 400 ug /gr of wet fungal tissue · Use cut tips for pipetting (not necessary for C. graminicola, but some fungi create a viscous supernatant for which a cut pipette will help) · … Continue reading
Fungal cultures preparation for maxi DNA extraction
1. Cut some mycelial plugs (like 5) from a 10 -14 days old PDA culture. 2. Place the agar plugs in 250 mL Erlenmeyer flask with 40 ml of Fries medium. 3. Add 5.2 microliters of HYGRO to each flask. … Continue reading
Anthracnose Leaf Blight Assay
Maize plants are typically grown the V-4 stage (about 3 weeks old) in Cone-tainers (small narrow cone shaped pots). Plant Inoculation Assay Potting the plants 1. Corn seeds are planted in soil that is 2:2:1 College Station field soil: Metro … Continue reading