Hi everyone,
I am a new PhD student with very little microbiology experience and I have a few questions regarding the growth of Lactobacillus Reuteri which I hope you can help me with.
I want to grow L. Reuteri so that I can use it to create a standard curve for quantifying my bacterial qPCR data. I know lactobacilli are facultative anaerobes that grow best in anaerobic/microaerobic conditions. However, I do not have access to any anaerobic equipment in my lab.
I am planning to purchase the following Lactobacillus reuteri Kandler et al. (ATCC® 23272™) strain from ATCC which says it is aerobic on the product webpage so I am assuming I can grow it aerobically? (https://www.lgcstandards-atcc.org/products/all/23272.aspx#generalinformation).
Questions:
1. Could you please provide me with a step-by-step protocol with the best way to grow this ATCC® 23272™ strain in the lab without anaeorobic equipment?
Since I am not interested in the fermentation products, I only need to make a standard curve so surely aerobic growth should be fine?
2. If not, could someone provide me with a step by step protocol for microaerobic growth without specialist equipment?
3. From your experience, which lactobacillus species grow best in oxygen?
I have a general idea on how to grow it but I am really struggling without access to any protocols or any Microbiologist contacts... The research papers I have found are too vague. I would really appreciate your advice.
Best wishes
Felicity Kate
I think this is the blind (me) leading a severe myopic,(you) sorry;-)
I have not helped write anything on culturing bacteria in centuries
if I remember correctly you need to anaerobically culture first
so anaerobic chamber and then DeMan Rogosa Sharpe Broth. Once you have a sufficient quantity of lactobacillus reuteri then you can test it against bacteria on agar in the open air or a cleanroom. My engineering students had to devise cleanrooms as a project and the results were not nightmares. What are the objectives of your experiments and that should dictate better the procedure, like where will get your bacteria, milk cheese, maybe saliva etc and so on
Hope this is at least a start.
Good Luck
Chris
Hi
This my work in the growth of Lactobacillus reuteri . Please see the attach files
Dear Felicity Kate
I have earlier worked with lactobacillus species where I used to grow them with traditional method as my lab had no equipments for growing anaerobes. I used to use candle and jar method, jar was a typical bell jar (pictures attached). The steps are as follows:
i) First streak or spread the bacteria in your desired agar or broth (I used MRS agar/broth)
ii) Stack them in a plain surface
iii) Light a candle
iv) Cover it with the bell jar
v) Wait until the candle goes off
vii) You will observe your bacteria growing after 24-48 hours
This method creates a microaerophilic condition. However, the conditions are not 100% anaerobic. All the best with your work.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology