Materials and Methods
Students were prepared for the exercise by reading assignments of literature on hairy root cultures. The literature included three research journal articles and an overview of the process in their lab manual. The instructor gave a short lecture on A. rhizogenes and plant root transformation. Reading, lectures, and student discussion on the hairy root transformation system occurred prior to the laboratory exercise. Due to time limitations, the instructor planted the soybean seed and prepared bacterial cultures for the class.
Materials that are needed for transformation:
Soybean seedlings—one 6‖ pot can supply enough cotyledons for an entire lab section.
24 hour A. rhizogenes cultures (containing a binary plasmid of interest)
Sterile Whatman filter paper
Sterile Paper towels
Sterile 1/4 strength Murashige and Skoog (MS) salts (no sucrose) or 10mMMgSO4.
Micropipettes
70% ethanol
Sterile surgical steel blades