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Author Topic: micropropigation preparation workspace  (Read 12213 times)

ONandONandON

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micropropigation preparation workspace
« on: January 05, 2019, 10:57:52 pm »

Greetings STS, nothing to post about yet, hopefully eventually, still in the gathering supplies and researching stage.
So i thought i'd invite everyone to join in! Feel free to post your questions, videos, experiments, helpful information..

Also trying to raise any interest in a group buy?! of "Murashige and Skoog medium" that's MS medium for short..
That's the most commonly used growing medium in micropropigation suitable for a wide range of plants.example:

https://en.wikipedia.org/wiki/Murashige_and_Skoog_medium
There are 22 chemicals needed, in mg amounts, each can be purchased in large amounts cheap..
i was thinking if people are interested, we could each purchase some of the chemicals, then trade.
Even if only two people, it would make it half-price, and the more people the cheaper it would be.

So post here if interested in the "Murashige and Skoog medium" group buy  ;D


« Last Edit: January 05, 2019, 10:59:40 pm by ONandONandON »
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DarkPines

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Re: micropropigation preparation workspace
« Reply #1 on: January 06, 2019, 01:20:27 am »

Dude, OMG I am so down for this!!

Shamichael

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Re: micropropigation preparation workspace
« Reply #2 on: January 06, 2019, 06:36:07 am »

Count me in.
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Neebu

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Re: micropropigation preparation workspace
« Reply #3 on: January 06, 2019, 12:05:16 pm »

An interesting project, but unfortunately my financial situation is not that good at the moment.
Nevertheless I would be very happy if you could keep us updated on your progress :)

I can offer 2 PDFs which might be interesting for doing experiments with tissue culture, but I´m not sure if it is allowed to post the links here:
Hirenkumar Sherathiya - Practical manual for Plant Tissue Culture: Basic Techniques of Plant Tissue Culture and Molecular Biology
Roberta H. Smith - Plant Tissue Culture: Techniques and Experiments

Edit:
Why don´t you just buy the MS Medium as dehydrated powder for making a solution, wouldn´t that be easier?

Edit 2:
I looked a bit closer, powder for 10L solution is not that expensive, depends on how much solution is needed?

« Last Edit: January 06, 2019, 01:50:12 pm by Neebu »
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DarkPines

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Re: micropropigation preparation workspace
« Reply #4 on: January 06, 2019, 06:12:04 pm »

Yea Im down to just buy my own mixture and experiment with plants I have here right now. Id love to share results, and try experiments with other people. Can any of this be applied to fungi possibly?

ONandONandON

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Re: micropropigation preparation workspace
« Reply #5 on: January 06, 2019, 07:32:31 pm »

On second look at prices, it would be MUCH cheaper and easier to buy it premixed  :P
i found some from this guy in Germany on ebay: 100L for 55$ 50L for 36 10L for 20
https://www.ebay.com/itm/Murashige-Skoog-Medium-mod-with-Gamborg-Vitamins-powder-premix-for-50-L/113205340598?hash=item1a5b9069b6:g:7wIAAOSwel5bjO-V:rk:5:pf:0
i guess i was only looking at the USA sellers, but thanks for the suggestion Neebu!

Awesome to see others interested in micropropigation! i'll be getting the 50L or 100L soon as i can..

so the group buy of individual chemicals is off.. but i guess we could have one for the premixed 100L ..
i only got 10$ paypal ATM, but i have ebay store selling plants, so not to long and i'll have more funds.
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ONandONandON

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Re: micropropigation preparation workspace
« Reply #6 on: January 06, 2019, 07:54:24 pm »

Can any of this be applied to fungi possibly?


Of course, it's very similar techniques and equipment needed for both.
 
-sterilizer / pressure cooker
-still air glovebox or hepa flowhood
-growing containers
-chemicals for cleaning

that's most of the basic equipment.

There is a ton of how-to videos on youtube..
Just search micropropigation or tissue culture.
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Neebu

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Re: micropropigation preparation workspace
« Reply #7 on: January 06, 2019, 08:23:42 pm »

I found the same product by dephyte, but on another site (it´s not cheaper there).
If you don´t mind, I would like to participate with 15$ for a bit of powder, this seems affordable.
I don´t have any other Equipment yet, but hope to start something some day  ;D

I attached the product datasheet.

Thanks for posting that thread ONandONandON.

Wish you good luck and all the best for your experiments!
« Last Edit: January 06, 2019, 08:28:44 pm by Neebu »
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ONandONandON

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Re: micropropigation preparation workspace
« Reply #8 on: January 07, 2019, 08:26:34 pm »

Awesome!  ;D
that's two people in for 15$, so we're at 10L for 20$ at least (but surely someone else will get in on this)

Let's try for 100L (it's the best deal) one more person makes 18.66$ each..
Two more (total of four people) is 14$even, each.. that's 25L worth each..
So two more people in would be perfect, and easy to divide into 25% each.

56$ https://www.ebay.com/itm/Murashige-Skoog-MS-Medium-for-plant-micropropagation-powder-for-100-L/123309579751?hash=item1cb5d2dde7:g:G90AAOSwt0dcEAcy:rk:11:pf:0

Edit: Or if shamichael and darkpines is in for 14$, that makes four and this groupbuy is closed, going onto next stage.
« Last Edit: January 07, 2019, 08:36:32 pm by ONandONandON »
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Neebu

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Re: micropropigation preparation workspace
« Reply #9 on: January 07, 2019, 09:17:39 pm »

It seems like, besides sugar and agar, that also at least an auxin is needed for the preparation of the MS medium.
In the books NAA is suggested for callus and root growth induction and it seems to be the cheapest choice (not sure if it is really necessary to add a cytokinine as well?).
I looked on the internet a bit and found a company which offers 25g of NAA for around 29 USD, shipping included.
I´m not sure if they will sell it to individuals, but buying from ebay might be an alternative.
I could take care of this (but only next month), so you don´t have to bear all the risk.

Also not sure if some kind of antifungal/antibacterial additive is needed?
Afair many antifungal/antibacterial substances are toxic to the explant.
Although Sherathiya writes that he uses an antibiotic called Cefotaxime, idk where to get something like this, which can be used for tissue culture and is not toxic for the explant.
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ONandONandON

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Re: micropropigation preparation workspace
« Reply #10 on: January 07, 2019, 10:03:48 pm »

Thanks for posting that product data sheet BTW! very useful.. anyways...
i think it's optional to add hormones and/or antibiotics, but might be needed for the best results.
The MS-medium already has some growth hormones (for example Indole Acetic Acid 1-30 mg/l and Kinetin 0.04-10 mg/l)
(i believe cytokins and auxins can be used alone or together in different concentrations to get different effects)
i recently bought some hormones to experiment with from ebay.. and might can trade/spare some of:
6-BA 6-Benzylaminopurine 99% 5g CYTOKININ type
6-KT Kinetin Furfurylaminopurine 99% 10g natural CYTOKININ type
IBA Indole-3-butyric acid Indole butyric acid 98% 10g Water soluable
Gibberellic Acid GA3 90% 5g
NAA Naphthaleneacetic acid Naphthalene acetic acid 98% 10g
DA-6 Diethyl Aminoethyl Hexanoate 98% 5g
Triacontanol 99% 5g

Also for cheap experiment trying out Antibiotic Neomycin Sulfate Ointment 1 Oz Tubes
 (neomycin is similar in action to the common geneticin commonly used in tissue culture)
(will try water extracting the neomicin sulfate, also inactive-ingredients don't look harmful.)
Several antibiotics can be bought for farm-animal use, or for fish antibiotics petstore ebay etc.)

There are many types that can be tried and just need some researching, here is some good information:
https://www.sigmaaldrich.com/life-science/core-bioreagents/learning-center/antibiotic-selection.html
https://www.sigmaaldrich.com/technical-documents/protocols/biology/antibiotics.html
https://www.sigmaaldrich.com/technical-documents/articles/biology/antibiotic-kill-curve.html

Following reagents are not included and need to be purchased separately:

    Distilled water
    Gelling agent, e.g. Agar or Gellan (if a solid media is required)
    Sucrose (20 - 30 g/l, depending on the culture requirements)
    Plant Growth Regulators (if necessary)

@lso Darkpines and Shamichael please confirm if you are in or out, thanks :)
« Last Edit: January 07, 2019, 10:12:28 pm by ONandONandON »
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Neebu

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Re: micropropigation preparation workspace
« Reply #11 on: January 07, 2019, 10:46:34 pm »

ONandONandON, you seem to be really well prepared in regard of plant growth hormones  ;D
I still need to do much more research on this topic.
Many thanks for the links and additional information!
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DarkPines

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Re: micropropigation preparation workspace
« Reply #12 on: January 08, 2019, 11:04:12 pm »

I can throw down about 20$ for this! My only problem is how to pay for it? If we are using ebay, I can go buy a gift card and just load up the code when needed. Let me know! Ill go buy one in the next few days.

ONandONandON

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Re: micropropigation preparation workspace
« Reply #13 on: January 09, 2019, 09:12:42 pm »

Ok cool, i currently have 24$papal, and two sales 'awaiting payment' so i think we got enough!
it's better if you have paypal and can just send it to me that way, and i will make the purchase.

(One of you hopefully has a paypal to send, or this will take longer)
IF not, the best way i know of is send me cash in a card in the mail,
and i should have enough paypal by then to cover online purchase.
Or, to use an ebay card, i can post a special listing.. loses $ to fees.

(56$100L)14$each=25% =25L each, i'll also send some hormones..
i estimate shipping about 3-5$each? though it's hard to say exactly,
it doesn't say how many grams we will be getting, just 100L worth.

« Last Edit: January 09, 2019, 09:14:33 pm by ONandONandON »
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DarkPines

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Re: micropropigation preparation workspace
« Reply #14 on: January 09, 2019, 11:23:14 pm »

Ok cool, i currently have 24$papal, and two sales 'awaiting payment' so i think we got enough!
it's better if you have paypal and can just send it to me that way, and i will make the purchase.

(One of you hopefully has a paypal to send, or this will take longer)
IF not, the best way i know of is send me cash in a card in the mail,
and i should have enough paypal by then to cover online purchase.
Or, to use an ebay card, i can post a special listing.. loses $ to fees.

(56$100L)14$each=25% =25L each, i'll also send some hormones..
i estimate shipping about 3-5$each? though it's hard to say exactly,
it doesn't say how many grams we will be getting, just 100L worth.

Yea thats my dilemma lol. I dont have paypal, and all my information is in an account I cant access because of forgotten security questions/answers. I might be able to access it if I call them maybe or email or something. Ill have to check into it in the future. I guess I can send cash, I have some trusty envelopes for it. Shipping would only be about 4$ or so. Not a big deal for this experiment. I lack all knowledge of this area and want to learn more by just kind of diving straight in to it lol.
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