Murashige and Skoog media preparation
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  #1  
Old 07-18-2017, 02:10 PM
cfreidsma cfreidsma is offline
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Murashige and Skoog media preparation
Question Murashige and Skoog media preparation

I got this today, and it didn't arrive with instructions. I am assuming that I follow the instructions for agar, and add this to the agar before it cools. I just want to make sure that I don't need anything else.

HiMedia PT022-1L Murashige and Skoog Shoot Multiplication Medium A with CaCl2, Vitamins, Sucrose, IAA, Adenine Sulphate and 2iP, 1 L: Amazon.com: Industrial & Scientific

I am planning on doing tissue culture of assorted plants, orchids included possibly.

Edit:
I am going to add sucrose, benzylaminopurine, naphthalene acetic acid, and the agar.

Last edited by cfreidsma; 07-18-2017 at 06:15 PM..
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Old 07-19-2017, 11:37 AM
dansyr dansyr is offline
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Short version, prepare it all together in one bottle at room temperature, non-sterile. The agarose won't dissolve at RT and that's ok, when you sterilize the media the agarose will melt and dissolve. The goal is to make sure the MS powder is mostly dissolved. If you can, pH adjust to 5.6-5.7. If you're in the kitchen, it might be a good idea to microwave until the agar melts so you can aliquot your media into final vessels and then pressure cook/autoclave. Usually, since I live in a lab and it's easy to maintain sterility, I'm lazy and just autoclave the media bottle(s) and aliquot afterwards (so I can skip the microwave step). And you can definitely do this too, but if you're in the kitchen, every time you open anything the odds of contamination are much higher so the microwave step might be worth it.

If you're in a lab setting, the optimal way to prepare media with heat-sensitive additives is to get media without those additives, (e.g. plain MS + vit) and filter-sterilize concentrated solutions of whatever additives you want. So you would be able to prepare and autoclave MS + vit + agar, let it cool a bit, then sterilely add whatever small volumes (because you made concentrated stock solutions) of hormones and C sources, mix, and then pour your plates. But that assumes access to lab stuff.

Sorry that got long winded. But media prep is basically all I do lately so I thought I'd share
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Old 07-19-2017, 01:42 PM
cfreidsma cfreidsma is offline
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Thanks, that helped quite a bit

This is my first time trying anything like this, and I don't have access to a lab. Wish I did though.
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Old 07-20-2017, 01:25 AM
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You may have a community college nearby with a microbiology lab. It might be worth your time to take a course. Often once you know the instructor, and explain what you are going to be doing, they may allow you to use the lab facilities for your own project.
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