Does anyone have any experience with this? Made liquid media first autoclaved and later added agar to same liquid media and did second autoclave for agar addition. Any such experience with pea broth media or any other?
Muhammad Shahid Iqbal woud it make any difference if I do 2nd autoclave adding agar?
Autoclaving aim at sterelisze and solubilize the constituents of the culture medium.
I dont think, it is necessary to do that; By doing that, you are waisting energy and time
F. Romain Fouelefack I acknowledge that. I just wanted to ask as I was running out of time to make agar pea broth nutrient medium and I just added agar in the liquid medium I had. So wanted to know if it makes a difference or not in prior. However, I am going to find the answer when I see the growth of the strains that I will culture and will update. But for now, I was seeking if anyone had prior experience with it.
I dont have any experience in that but, I think no change will be observed
Its harmful for media components. Some time it could spoil functions of PGRs.
Dear Dr.,
There is no problem with agar using in the second time, but you must notice that you do not know anything about fertilizers concentration and hormones, beacause you have had added fertilizers to agar in the first time.I hope my comments come into your hands.
Best regards,
Avestan
Yes I did this before and I didn't notice any problems with Agar. Only probably lose a little bit of solidity. I also added nutrients
Dear Bikal dai, I have done it in PDA , But I haven't noticed any difference between single autoclaved media and double autoclaved media ......i have done this during isolation of fusarium fujikuroi
Sorry for the missunderstanding. Did you mean about PDA media for microbial culture? I answered before for tissue culture MS media. For tissue culture media its harmful for double autoclaving. But for PDA, there will be potato starch, dextrose and agar for silidifying. So it could be ok but i never did this.
If you wants to add agar , add it at the time of preparing your media. first dissolve your media powder and than add agar. you need to dissolve the agar properly by heating in oven so that no lumps are formed. after that you can autoclave it. there is no point in adding agar after autoclaving and than again autoclaving.
I'm worried with the hormone and vitamin, if we re-autoclave the medium. I'm thinking about two times in heating. If that the only choice it's ok, but I don't want to give the recommendation in other way, Ara-san. Ganbatte for your research ^-^
Actually Bella, for PTC media with PGRS it will be harmful because chemical bonds will be broke. But this question may be not for PTC media, author has been asked for PDA media. For PDA i dont know, it will be ok or not. But double autoclaving of any nutrient media could be harmful.
In case of PDA media it will not be a problem. But i think you need not do the twice autoclave. In the liquid media you should add the solidifying agent before autoclave.
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