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Thread: plant tissue culture M&S Agar

  1. #1 plant tissue culture M&S Agar 
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    I made M&S agar but it has not set, i would like to know what I did wrong

    Recipe
    0.8% agar
    4.605g/1000ml M&S
    3% sucrose
    3mM MES (1ml 0.9M in 300ml)

    autoclave

    so to 1litre water i added 4.60gg M&S and stirred gently, then I added 8 grams agar and stirred and 30 grams sucrose and stirred. in 3.3 mls water I added 0.578g MES. I stirred then autoclaved. This morning however when cooled they are a yellowish colour and liquid.

    What did I do wrong?


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  3. #2  
    Forum Professor Zwolver's Avatar
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    i don't know the agar.

    I would ask myself the following questions.

    1) Has something grown inside it??
    2) Was the pH right??
    3) Did i add enough agar??
    4) Weren't some chemicals to be added AFTER autoclaving due a sterile filter??
    5) Were the ingredients out of date??
    6) Did the autoclave have the right temperature and time??

    all right... then i don't know..


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  4. #3  
    Forum Freshman Tara's Avatar
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    Quote Originally Posted by Zwolver View Post
    i don't know the agar.

    I would ask myself the following questions.

    1) Has something grown inside it??
    2) Was the pH right??
    3) Did i add enough agar??
    4) Weren't some chemicals to be added AFTER autoclaving due a sterile filter??
    5) Were the ingredients out of date??
    6) Did the autoclave have the right temperature and time??
    Plus sometimes kind of agar is important, special about mark.

    Sincerely yours
    Tara
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  5. #4  
    Forum Freshman shazzy's Avatar
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    some agar did not solidify becoz of the PH
    Tara likes this.
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  6. #5  
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    Hi guys!I'm a newbie here.

    Regarding M&S, can i use the macro, micro , iron sources, vitamins stock solutions that have been prepared about 6 months ago?

    TQVM!
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  7. #6  
    Forum Freshman Tara's Avatar
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    Quote Originally Posted by nondagreat View Post
    Hi guys!I'm a newbie here.

    Regarding M&S, can i use the macro, micro , iron sources, vitamins stock solutions that have been prepared about 6 months ago?

    TQVM!
    Hello, welcome to forum,
    As I know every stock just have to be for maximum 1 month, so definitely stocks that 6 months passed on them is not really good. Because f evaporating, loosing their characters (material I mean espesial Iron), also pericipitation of materials in the stocks... There are some more reasons too for.

    Regards
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  8. #7  
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    Bacteria food has to be free of bacteria to start with.What do they eat mostly sugar?I've heard keeping it keeping it refrigerated helps.Nice fresh food for the bacteria to eat and grow,
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  9. #8  
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    Also, I had this problem before when I poured my first agar plates. The agar appeared melted, but I put the autoclave on the shorter setting (out of laziness) and consequently, the agar had dissolved, but not melted (while the solution was uniform, it was not clear).

    3 suggestions:
    -Autoclave longer
    -Be sure to swirl very well after autoclaving (agar settles)
    -Make sure the solution is clear, not cloudy

    Good luck!
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  10. #9  
    Forum Freshman Tara's Avatar
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    Quote Originally Posted by iRNAbooks.weebly.com View Post
    Also, I had this problem before when I poured my first agar plates. The agar appeared melted, but I put the autoclave on the shorter setting (out of laziness) and consequently, the agar had dissolved, but not melted (while the solution was uniform, it was not clear).

    3 suggestions:
    -Autoclave longer
    -Be sure to swirl very well after autoclaving (agar settles)
    -Make sure the solution is clear, not cloudy

    Good luck!
    Hello, Good idea but still here is a thing, if we use longer time autoclove it might also disturbs matrials, as hormones or breaking their sugars and something like that.. I think using better marks for materails is the best solution.

    Regards
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