Hi . I have moraxella isolates in tryptose soya agar slant for period one month but i do subculture in brain heart and tryptose soy broth but no growth . what can i do for maintain for some months?
I will prefer to go with large size bacterial inoculum for preservation. The chances of bacterial death during and after preservation are evident so we need to take few more steps to get viability after preservation. First as mentioned, try to take large size of inoculum, so that some bacteria will revive even after death during preservation. Second, do not give direct shock to bacteria, by keeping it in -80 degree C. instead give them step wise shock, by first keeping in -4 for few hours and gradually decreasing the temperature. This can also reduce killing of bacteria during preservation. The rest is same for shorter duration, you can go for preservation in Glycerol, while for longer duration, you should try lyophilisation. I hope this will help.
first make a massive culture onto petri dish with Mueller Hinton Agar. Then, you harvest it with 15 ml of sterilized Skin Milk 20% w/v, and dispense the bacterial suspension into cryovials and preserv to -70°C.
You can use either TSB +15-20% glycerol or BHI +15-20% glycerol or skimmed milk as mentioned above. If you store in -20C, it'd better do subculture the bacterium every 2 months.