Author Archives: Mike

THE BEST METHOD WE HAVE FOUND (modify from Lisa Vaillancourt Lab) (Method of John Tuite, modified by Rick Latin, 9-1-95) 1. Fill vials  (cryovials Wheaton 8 ml, cat 224884 we get them from VWR) half full with silica gel granules, … Continue reading →

(from Slavica)   5 minutes 70% ethanol. 1:45 hours 10% H2O2   In a 50 mL tube, rinse seeds with sterile water 4-5 times.   If seeds will be used for In vitro tissue culture: Seeds will float on H2O2, … Continue reading →

                                                                                                                  (Modification from Lisa Vaillancourt Lab)    1. We use TOP10F’ cells (Electrocomp Kits, INVITROGEN cat C6665-55, Is in the -80C. READ CAREFULL Kit information, print … Continue reading →

  1. Harvest conidia from 2 or more plates that are 3-4 weeks old, center inoculated. Expect about 1×108 conidia per plate. 2. Wash conidia once with sterile water. Resuspend the last time in Fries medium. 3. Count with hemacytometer. … Continue reading →

1. Colletotrichum graminicola is grown on Petri dishes of Potato Dextrose Agar (PDA) at room temperature under fluorescent lights. The cultures should be 12-15 days old. 2. Place 8-10 mL of sterile distilled water on the surface of the Petri … Continue reading →

From Lisa Vaillancourt Lab   1. Prepare PEG solution and place on a rocker to mix. 2. Thaw protoplasts on ice. 3. Place 100 mL (10^7 protoplasts) in a pre chilled 50 mL centrifuge tube. 4. Add 3 ug of … Continue reading →

  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 →

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 →

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 →

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 →