I need to grow my bacteria which is Streptomyces sp. from spore suspension method. I have grown them onto the nutrient agar by spread plate method and now I incubate them for 5 days. And for the next step, I will prepare the stock from the spore by scrapping the spores with sterile saline Tween 80 solution and mix them with glycerol solution as well as transfer 1 mL to eppendorf tube and stored in -4C for further used.
My question is if i want to inoculate them to production medium which is MSM, do I need to activate back the spore stock in nutrient broth, incubate for 2 days, then homogenize them by using waring blender and check the OD & inoculate them to the medium or can I just transfer 1 mL of the spore stock directly to MSM without activate them and homogenize by using blender?
I really need the explanation on this. Thank you
It is normally accepted that before the production stage, a fresh inoculant should be prepared. Threfore, you should refresh the spore material...
Hi,
Prof. Nikolay is spot on. Though some researchers would directly inoculate the spores into the media, it is better to do a pre-culture, and then use it as the starter inocula. FOr my experiments, both the approaches have worked.
It really depends how fast they grow, if they grow very fast then use them if they grow very slow then the activation step is your only option. Bear in mind that the purpose of storing spores in glycerol in -80C or as mentioned in ur post is for use when something wrong happened to ur daily usage cells (died or contamination ....etc), hope this helps.
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