Question

list all of the steps involved in sub culturing a colony from a plate to another TSA plate

Answer 1

Subculturing is the process of transferring microorganisms from one media type to another without contamination.

Trypticase soy agar or tryptone soya agar (TSA) are nonselective media that provide enough nutrients so that wide variety of microorganisms can grow.

Why subculturing is required?

Required to keep stock culture of organism for testing and also to see cultural characteristic

Steps involved in sub culturing a colony from a plate to another TSA plate:

1) Place the tube which has culture in the space between the ring and middle fingers of the left hand.

2) Place the new tube that is to be inoculated in space between middle finger and pointer finger which is parallel to the old tube. Hold these tubes away from body.

3) Loosen both the caps

4) Hold the loop or needle with the right hand.

5) Flame the instrument. Flaming the tubes immediately after uncapping and before recapping is necessary as if there are microbes the tube then these may come into the nutrient base, flaming will not allow this.

6) Cooling and remove caps from both of the tubes. Cooling the inoculating instrument prior to obtaining the inoculum as excess heat can fixate the samples or kill the organisms.

7) Remove caps and be ensure that both caps should be removed in only one motion.

8) Flame the opening of tubes.

9) Insert the instrument into the old culture and then immediately transfer to the new tube.

10) Flame the tubes. Flaming the inoculating instrument prior to and after each inoculation is required to maintain sterility.

11) Label the glass tubes