Questions about growing Streptococcus Mutans
Posted: Tue Jan 11, 2022 3:42 pm
Hello! I have a couple of questions I am planning on ordering a live culture from Ward's (Link below - I am ordering the tube with the live bacteria, not the lyophilized one). I want to grow the culture in an MSAT Agar plate for my experiment. My experiment is about testing copper water's effect on the oral microbiome. I have two questions:
1. Would my procedure for inoculating the agar work? I put it at the bottom of the post.
2. I am planning on inoculating the agar once to practice. To do that, I would need to open the culture vial and remove some of it to inoculate. For the actual experiment, can I use the opened vial again? Do I need to store it in low temperatures to use it again? If I cannot use the vial again after opening it, I was planning on removing some of the bacteria from the practice agar plate and subculturing it into another plate for the actual experiment. How would I store the practice agar plate that has already been inoculated so I can remove some of the bacteria and reuse it for the actual experiment?
One more question! Can I post my procedure here with a bunch of questions I have about it and hopefully get some advice on it? I know it's a big ask, but I would really appreciate it if that was possible.
Thanks a bunch in advance!
Link for Streptococcus Mutans:
https://www.wardsci.com/store/catalog/p ... .jsp.order
Procedure for inoculating Streptococcus Mutans:
a. Sterilize half a cup of water in the microwave for 1 minute. Wait for it to cool to room temperature.
b. Light a Bunsen Burner.
c. Put on nitrile gloves for the following steps.
d. Open the test tube of live Streptococcus Mutans and pass the opening through the flame 3 times. Wait for it to cool.
e. Take a sterile pipette and draw up 0.5 ml of the culture from the test tube. Remove the lid of an agar plate and place 0.2 ml into it.
f. Take a sterile cotton swab and dip it into the sterilized water. Press the cotton head against the edge of the container and apply pressure to get rid of excess water. Hold the pipette in another hand while doing this. Do not let it touch any surface.
g. Streak the agar plate in one direction using the cotton swab to spread out the bacterial culture. Rotate the plate 90 degrees and do the same thing again. Repeat the process three more times, then dispose of the swab. Place the lid back onto the agar plate. Let it dry for 5 minutes.
h. Seal the agar plate using Parafilm.
I. Incubate the jar and the agar plate of S. Mutans in a 37 degree Celsius incubator for 48 hours.
1. Would my procedure for inoculating the agar work? I put it at the bottom of the post.
2. I am planning on inoculating the agar once to practice. To do that, I would need to open the culture vial and remove some of it to inoculate. For the actual experiment, can I use the opened vial again? Do I need to store it in low temperatures to use it again? If I cannot use the vial again after opening it, I was planning on removing some of the bacteria from the practice agar plate and subculturing it into another plate for the actual experiment. How would I store the practice agar plate that has already been inoculated so I can remove some of the bacteria and reuse it for the actual experiment?
One more question! Can I post my procedure here with a bunch of questions I have about it and hopefully get some advice on it? I know it's a big ask, but I would really appreciate it if that was possible.
Thanks a bunch in advance!
Link for Streptococcus Mutans:
https://www.wardsci.com/store/catalog/p ... .jsp.order
Procedure for inoculating Streptococcus Mutans:
a. Sterilize half a cup of water in the microwave for 1 minute. Wait for it to cool to room temperature.
b. Light a Bunsen Burner.
c. Put on nitrile gloves for the following steps.
d. Open the test tube of live Streptococcus Mutans and pass the opening through the flame 3 times. Wait for it to cool.
e. Take a sterile pipette and draw up 0.5 ml of the culture from the test tube. Remove the lid of an agar plate and place 0.2 ml into it.
f. Take a sterile cotton swab and dip it into the sterilized water. Press the cotton head against the edge of the container and apply pressure to get rid of excess water. Hold the pipette in another hand while doing this. Do not let it touch any surface.
g. Streak the agar plate in one direction using the cotton swab to spread out the bacterial culture. Rotate the plate 90 degrees and do the same thing again. Repeat the process three more times, then dispose of the swab. Place the lid back onto the agar plate. Let it dry for 5 minutes.
h. Seal the agar plate using Parafilm.
I. Incubate the jar and the agar plate of S. Mutans in a 37 degree Celsius incubator for 48 hours.