Transcript GMO Potatoes for Mentors

Bacterial identification
procedure
Part 1 : Preparation of
API miniaturized system
Materials avaliable
Physiological sterile
water tube (5 mL)
Distilled
water
Oil
Agar Petri dish
API system
with box and lid
Unsteril pipette
Sterile pipettes
1. Bacterial suspension
preparation
1.1. With a sterile inoculation loop, take a
bacterial colony on agar Petri dish.
WARNING! The sample is realized in asepsis
around the Bunzen burner.
1.2. Make a suspension with
this bacterial colony in a
steril tube of physiological
water (5mL).
In the water, rub inoculation
loop on the tube side to
disperse all the bacteria.
WARNING! The sample is realized in asepsis
around the Bunzen burner.
S
1.3
Homogenize your
tube by manual rotations
and write « S » on this
tube.
2. Preparation of the API
system
2.1. Fill up the bottom of the box with distilled
water to cover all the cavities.
2.2. Make a rotational movement with the box to
distribute the water in all the cavities. After that,
aspire the water excess with a unsterile pipette.
2.3. With a forceps, put the API system on the the
box.
2.4. Cover with lid and write your «table number»
on the plastic label of the box.
2. Filling the API system
Introduction
Cupule
Microtube
Examples :
Non underline
test
Underline test
Framed test
3.1. Open the box and use the lid such as a
support to place the box in an inclined position.
3.2 Homogenize your
suspension by manual
rotations.
Take 2 mL of the suspension
« S » with a sterile pipette.
S
WARNING! The sample is realized in asepsis
around the Bunzen burner.
3.3 Fill up all the tests with this suspension
respecting this next rules :
 For an underline test or not
(example : ONPG or ADH), fill
up only the microtube,
 For an framed test
(example CIT), fill up the
microtube and the cupule.
Cupule
microtube
3.4 With a sterile pipette, add oil in the 5 cupules
on the underlines tests (ADH, LDC, ODC, H2S,
URE).
Cupule
3.5 Close the box and incubate at 37°C during
16 hours.
API box
in incubator
4. Analyse your results
The second day, after incubation,
identify your bacteria (part 2).