** ph : 6.8-7.2

** memo :

Inoculate Rhizobium tumefaciens to TY liquid media (5g peptone, 3g yeast extract, 0.83g CaCl2?2H2O, L-1) and incubate at 30?C for 5 days by shaking with 100 rpm.

Centrifuge the cultured media at 8000 rpm for 20 min at 4?C and collect the supernatant not to uptake the bacterial pellet (Rhizobium tumefaciens)

Mix the cultured media and PY liquid media (10g peptone, 6g yeast extract, 4g NaCl, L-1) with 1 : 1 volume and adjust pH to be 7.2.

Add gellan gum to the mixed liquid media to be 1.5 % and autoclave.

Pour the media approximately 15 to 20 mL to each petri dish.

Inoculate novel species SA29B on the gellan gum media plate and incubate at 30?C for 3 to 5 days.

Remarks; It is required the caution because of that the bacterial colonies on the plate, which passed one week or more after colony formation, may not grow even inoculated on fresh media plate.

** additional actions :