How did you autoclave this large quantity of medium (20 L)? How much agar did you add? Looks like the agar did not completely melt.
And, what kind of medium is this? MS? The color did not look right (kind of brown).
This is the MS media with 7gr/l.each time a number of glasses are autoclaved.is not the reason why AGar isnt resolved? Or the reason it burns(Agar) when its boiling is tat the media isnt transparent.
There are a number of reasons why agar does not solidify. Judging by the color the most probable reason is overburning. You should check your autoclave to ensure the machine is not broken and temperature and time are correct. Also, the medium should be taken out soon after autoclaved and not left inside the autoclave for too long (swirling the medium when it's still liquid could help). Other reasons could be too low pH - check your pH meter - and bad quality of agar - boil some agar with water to check, maybe increase agar concentration.
Hi Parissa Mahmudi,
To your questions in the answer above.
1. We used 8 grams of agar per Litter of medium.
2. The color should not be 'brownish', like that in your picture. See the correct color of MS solid medium in the attachment.
"Why in the distribution method of culture media (large volume example 20 lit) before autoclave , the culture media isnt hardened?"
If the agar powder is pre-mixed with a large amount (20 L) of medium, and then distribute it into small tissue-culture containers for autoclave, then the agar powder in each container can be different. Some will have the amount than it requires, and some will have more agar than it requires. We prepare 1 litter medium containing agar powder in a big flask. After autoclave, the agar powder melt, then we swirl the flask a little to make sure the agar is evenly distribute in the medium. Then, we distribute it into tissue-culture containers in a sterile hood.
Hi. Dr
The red color of the medium is due to the replacement of a substance with another substance.in commercial works, its usually made in the form of a culture medium and the distribution of the medium after the autoclave is timely
I tested it yesterday and realized that it should be cooked(boiling) in two separate containers to be thoroughly disdolved.
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