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Help with fermentation please!!

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Hello Everyone


I am asking for a word of advice.


I am supposed to be doing fermentations soon. My problem is I never really had much experiences with these.



I need to run 3 fermentations like this.



1- Yeast DSM 1333 s cerevisiae


2- Yeast + Gram positive contaminant (Lactobacillus fermentum)


3- Yeast + Gram negative contaminant (E.coli)



I decided to use these conditions for 3 fermentations



Broth= Water 1L, yeast extract 10g/L, glucose 20g/L, peptone 20g/L


pH = 6


temperature 30C


160rpm



For starter cultures and yeast i used hemocytometer and the formula


initial concentration x volume i want to know = final concentration (start fermentation) x volume of fermentation (250mL).


so ill find what volume i should pour in to start fermentation


but i find very hard to count bacteria in hemocytomer. can I use OD600 to calculate the volume i need to pour of bacteria cultures>


if so how do I do it?



my bacteria (starter culture) ended growth in OD=2 can i dilute it? and sayOD = 1 = 10^7 cfu/ml?




Thank you for your time




Adriana


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