Ketolactose Test For Agrobacterium Biotype 1

OBJECT: To identify specific Agrobacterium strains.

MATERIALS:

MEDIA: yeast-extract indicator medium (YI).

REAGENTS: Benedict's reagent

EQUIPMENT AND SUPPLIES: Inoculating loop, Agrobacterium biotype 1 culture, Agrobacterium biotype 2 culture, Escherichia coli culture.


PROCEDURE:

1. With the help of waterproof marker or wax pencil, draw lines on the bottom of a YI plate to divide it into three pie-shaped sections of equal size.

2. Use flame-sterilized inoculation loop, streak one third of the plate with Agrobacterium biotype 1 cells, one third with biotype 2 cells and the last third with E.coli cells.

3. Mention your name, the type of cell in each section of the plate and the date on the bottom of the plate.

4. Incubate the plate in an inverted position for two days at 28-29 'c.

5. Flood the YI plate with 4 ml of Benedict's reagent.

6. Incubate the plate at room temperature for one hour.