Hello, you can use liquid media for grow culture and precipitate culture by

centrifugation and than you can prepare the culture suspension at which quantity you want. You can calculate prepared suspension CFU/ml by decimal dilutions in plates or spectrophotometrically by measuring OD at 600nm after creation relation between OD and CFU at 600nm.

Hope this info will help you.

Good luck !

Hello, you can use liquid media for grow culture and precipitate culture by

centrifugation and than you can prepare the culture suspension at which quantity you want. You can calculate prepared suspension CFU/ml by decimal dilutions in plates or spectrophotometrically by measuring OD at 600nm after creation relation between OD and CFU at 600nm.

Hope this info will help you.

Good luck !

You should not have problems on solid media, Bacillus subtlis growth is great unless you are using poor media. Can you describe the media you use and the incubation temp.

best wishes

Prepare different batchs to certain CFU that they are able to grow, Next centrifuge one of them and add it to another batchs in a way to get the proper CFU. Note that centrifuge them maximum in 5000g and not more than 5 min otherwise you will loose them

Let me put my question the other way

I am already getting 1 Billion cells per ml broth (1 x 10^9 CFU/ml) - Meaning need to get more cells per ml (more than inoculated CFU/ml)

Now, my objective is to produce a stock which contains 10 x 10^9 CFU/ml or 1 x 10^10 CFU/ml (approx)

Be it any bacteria just to give an instance, I have taken Bacillus subtlis (may be we could take some gram negative bacteria for example sake) here to explain. Also, I don't have the facility to lyophilize.

Your question is still vague. What do you mean by more than inoculated. If you have inoculated a certain number already and you desire to have higher concentration of cells, then you simply incubate the culture at appropriate conditions. After that you harvest and like Karen said, you centrifuge and regulate your starting diluent volume to prepare the needed stock suspension. Bacterial concentrations are such as could be prepared as the need arises. The more dilute your culture suspension is, the lower the concentration of cells. If you desire a large concentration then your stock suspension should not be too dilute. From that stock suspension, different lower concentrations can be prepared using appropriate diluent.

Hello, I think again problem with question explanation. If 10^9 you can receive without problem and what you finally need is 10^9 or 10^10 so what the question???

And what about gram negatives? It does not matter what cultivating if you have appropriate media and conditions. If you looking for how to increase from 10^9 to 10^10 it's already explained above.

Good luck.

Sorry for the confusion.

I am inoculating 10^9 CFU/ml - How to get higher concentrated 10^10 CFU/ml or even higher?

Now, hope I have clarified the ambiguity.

Thanks in advance.

Hi Aarya, I would apply a fed-batch strategy to obtain a high cell density

Regards

Juan Moreno-Cid What is this feed batch strategy? I tried searching but didn't get the exact working principle.

Interesting!