How To Make Plant Tissue Culture Media: The Ultimate Guide (Beginner, Intermediate, and Pro)

[@GSS_94]

95% success rate is great for a microwave. For beginners it's a good thing

[@PlantCellTechnology]

It’s a great way to get started!

[@fm00092]

I watched from beginning to end. It no longer feels impossible to do this as a beginner and test it out without spending thousands of dollars on equipment. Thank you so much!

[@PlantCellTechnology]

This is definitely achievable with a budget as low as $300. So, if you're eager to give it a try, go for it! And please, don't hesitate to reach out if you need any assistance along the way.

[@SunSolSys]

Thank you so much for making this video! I learned a lot from it and I will start watching through all of your videos next. This was a great contribution =)

[@PlantCellTechnology]

Great to know that you enjoyed your video!

[@mikearts6836]

Can you please clarify something? Its very confusing how you list. Add 1 ml of BA, Kinetin , or TDZ for multiplication media. Or NAA or IBA for rooting. Add 0.1 ml of NAA or IBA for multiplication media. OR BA , kinetin, or TDZ for rooting. Am i crazy or do these two comments contradict them selves? thank you!

[@aviationcompilation455]

Cant wait to take the online master class in May!

[@PlantCellTechnology]

Oh, you registered already!! Yay!! See you there!

[@spineless5990]

Another great video. Can you talk about disposal and recycling of ingredients and meterials please. Thanks.

[@PlantCellTechnology]

Absolutely! We have forwarded the idea to Francisco.

[@mindhunter00789]

very similar in a way to growing mushrooms sterility is key to succsess thanks for vid

[@PlantCellTechnology]

That's correct. Maintaining sterile environment is crucial for tissue culture successes.

[@wessamsaifeldin6715]

Realy a big big thanks for this informative video 💙💙💙

[@PlantCellTechnology]

Thanks for your love and support!

[@user-fw3zz5hh4i]

Can you please have video on how to add the explant into the media

[@DuaneEseo-ul1bg]

Thanks for sharing your technique.

[@PlantCellTechnology]

Hope you found it useful!

[@beryldiamond]

Ha! I used to do the “risky way” all the time pouring bacteria agar plates. I split the pile in half though, so it’s two piles of ten and not wobbly. 😃

[@beryldiamond]

Just finished the video and it was super informative! Thank you! I’ve done mammalian, bacterial, and intracellular bacterial cultures, but never plants. I’m going to be trying to start some micropropagation for my new plant business and possibly some in vitro germination. So it’s helpful to see the differences in making media and such. Thanks!!

[@prettyingreen]

Great vid, Francisco!

[@PlantCellTechnology]

Thank you! 🙏

[@antoniosanford4675]

Have you ever heard of 3dponics ? They've got an assortment of simple 3d printable pneumatic components that could be used to make bioreactors and biocouplers. I really want you to have a 3d printer.

[@antoniosanford4675]

Great video by the way!

[@PlantCellTechnology]

Happy to know that you found the videos insightful!@@antoniosanford4675

[@antoniosanford4675]

@@PlantCellTechnology when are you going to do a livestream?

[@PlantCellTechnology]

This is something to ponder on for us@@antoniosanford4675 ! We'll inform you whenever we plan to have one.

[@lukeblossom4870]

Am I the only one that notices the chapters are labled as: ingredients, chicken marinade, batter & mamas biscuits? 🤣🤣🤣

[@GSS_94]

Wait what 🤣

[@PlantCellTechnology]

What 😂😂

[@lukeblossom4870]

Its gone now but when the video was first posted it said that. The editors must be having fun :) @@PlantCellTechnology

[@PlantCellTechnology]

haha!!! @@lukeblossom4870

[@devonlockwood1477]

That's hilarious 😂❤

[@ghettosteeve]

Is there a way to make bulk media for storage without pouring into vessels immediately before or after sterilization? For instance, Can you sterilize liquid media without the gelling agent, store in media bottles, then add the agar later when pouring into your vessels? I’d imagine you would need to sterilize a second time in that case

[@PlantCellTechnology]

No, you can not do that. You need to sterilize the media after adding all the ingredients and use it instantly. That's how you ensure a complete aseptic environment for your plants. Because your media contains sugar, it's prone to microbial growth.

[@ghettosteeve]

@@PlantCellTechnology if I made too much media for one sterilization in my pressure cooker, would it be possible to store the media in vessels just long enough to finish my first sterilization? Or will I have to mix a new batch of media if it can’t fit.

[@PlantCellTechnology]

@@ghettosteeve Absolutely! You can do that.

[@anyadevine4094]

Hi I absolutely love these videos and I'm always excited for the next one to come out so i appreciate your hard work and effort. I'm currently trying to do some Tissue Culture myself at home however my PGRs are in powder form. Unfortunately, as I am in the UK this was the only PRGs I could find. Please could you do a video on how i can turn these into a solution? Thank you! P.S. My NAA powder is 98% and I'm not sure what this means in terms of diluting it.

[@PlantCellTechnology]

Hi, We do have video on the subject on our channel, please check it out: kzbin.info/www/bejne/jnqWoXuVjcRniqs

[@anyadevine4094]

@@PlantCellTechnology Amazing thank you so much!

[@josephbevilacqua6126]

Great video as always! Question about the Gamborg vitamins: do you typically add in place of standard MS vitamins or do you add the Gamborg vitamins in addition to the MS vitamins?

[@PlantCellTechnology]

Gamborg media and MS media are two different media with different macro and micro nutrient compositions. They are used for the tissue culture of different plants based on plants' requirements.

[@surajsagar6935]

I want tisue culthare orchid plant can u help me frd

[@nunosousa9767]

Thank you for this content - Great video! Do you dispatch the items available on your website to the UK?

[@PlantCellTechnology]

Yes, we do! We ship worldwide. What do you need?

[@fatmatunaboylu8122]

Can I use lime juice instead of HCL or vinegar to lower pH? Because I use lemon juice safely. I also have many baby orchid production videos on my channel. Bayy.

[@rajnikantpopat4666]

Can u not purchase prepared media ? If yes. Where from to purchase? If not able to purchase, what is the reason?

[@PlantCellTechnology]

Of course, you can purchase prepared media. Like you will just need to weigh and dissolve the powder in distill water and autoclave it for use. Here's the link to the MS media: www.plantcelltechnology.com/gelling-agents/

[@muddypawswanderlust]

I did everything like what you did in the professional tutorial but I'm getting way too much condensation and all my plants are exhibiting vitrification (hyperhidricity)! Im so frustrated 😫 how can I prevent this?

[@muddypawswanderlust]

My media is based on DKW and my gelling agent is agar (7gr/L) and after pouring my media, I kept the lids open for 10minutes and after they solidified, I closed the lids but after 48hours there is too much water on the surface of the solidified medium

[@PlantCellTechnology]

Read this article to learn about the ways to remove vitrification from your cultures: labassociates.com/hyperhydricity-in-tissue-culture-and-methods-to-deal-with-it Three things you need to focus on are the temperature and light you provide plants and type and amount of gelling agent.

[@PlantCellTechnology]

Avoid keeping the plants very close to LED lights as well @@muddypawswanderlust

[@muddypawswanderlust]

@@PlantCellTechnology thanks 🙏

[@PlantCellTechnology]

Happy to help!

[@succulenthunter9916]

Do you sell all the media, vitamins and growth regulators?

[@PlantCellTechnology]

Yes. You can purchase all your tissue culture requirements from our store: www.plantcelltechnology.com/pct-store/

[@valeriarobles1761]

Hi Francisco thnks for the video :) could u teach us how to prepare the 1 M NaOH/HCl Solution! SAludos :)

[@PlantCellTechnology]

Hi, We already have a video on that on our channel. Please check out this video: www.youtube.com/@PlantCellTechnology/search?query=stock

[@HappyDayRoots]

Another huge thank you

[@PlantCellTechnology]

Thank you for watching!

[@m3rky240]

Do you guys do live classes in PA?

[@PlantCellTechnology]

PA as in?

[@m3rky240]

@@PlantCellTechnologysorry for not clarifying this. I meant Pennsylvania

[@PlantCellTechnology]

@@m3rky240 We do live classes that you can join from any part of the world.

[@danieljheelan5256]

Hi Francisco, Want a video on the products needed and how to create plants using this method. Daniel

[@PlantCellTechnology]

Hi Daniel, Please check out other videos on our channel to get an idea of the applications of tissue culture and the required chemicals and equipment. And, if you want to learn more you can check out our blogs curated to help culturists like you in the journey: www.plantcelltechnology.com/blog/ You can even send your questions related to any blog to our scientific writer, Anajli, who will help you with your questions. We hope these resources help you at the best. We're creating many more educational videos, so stay tuned to our channel.

[@danieljheelan5256]

@@PlantCellTechnology Thanks a lot. Daniel

[@adedoyinadetunji2078]

When it comes to media sterilization, i have seen some laboratory using filter paper to filter their media. Is this truly necessary of normal autoclaving will do the work? Is there an added advantage to using vacuum filtering apparatus? Thank you.

[@PlantCellTechnology]

Not all plant hormones are autoclavable. Some degrade at higher temperatures, such as zeatin and kinetin. Therefore, for these hormones, a filter sterilization technique is used and they are added to the media after the media is sterilized. However, you can autoclave other hormones, such as BAP and NAA, with the media.

[@JiaMaruji]

❤❤❤❤❤

[@elizabethrodarte3986]

When preparing media from stock solutions, is it necesary to add the vitamin solution after autoclavimg the media?

[@PlantCellTechnology]

Which stock solution are you talking about?

[@elizabethrodarte3986]

@@PlantCellTechnology MS stock solutions. Macros, micros.. One of them is the vitamins solution and just wanted to know if i can add them before autoclaving like when using the powder ms media that already includes vitamins (cause it'll make things a lot easier and faster) or if its necessary to add them after autoclaving the media like ga3.

[@PlantCellTechnology]

Yes, you need to autoclave the MS media with out without vitamins. @@elizabethrodarte3986

[@marvin216]

Hey there, I am currently following a protocol where hormones are added after sterilization, does this make a difference? Are the hormones temperature sensitive?

[@PlantCellTechnology]

A few hormones are. It depends on what hormones you are working with. Synthetic hormones can generally be autoclaved with the media. However, natural ones are preferred to be added after autoclaving the media through a the filter sterilization technique.

[@marvin216]

@@PlantCellTechnology The protocol uses TDZ, GA3 and IBA, they all should be autoclaveable right?

[@PlantCellTechnology]

@@marvin216 No. GA3 and TDZ should be filter sterilized, and IBA can be autoclaved.

[@marvin216]

@@PlantCellTechnology thank you sir! What workflow do you suggest to store and add the non-autoclavable hormones? A) Dissolve the hormones in water/buffer stock solution, then sterile filtering the hole stock solution and keep it sterile (only open in clean bench) B) Dissolve the hormones, keep them non-sterile and sterile-filtering only small batches (1-2mL) as needed. C) combination of A)+B): Sterile filtering the whole solution in the beginning after dissolving the hormone, then also filtering again each small batch of the solution that is being used. I hope it is clear what I mean, I fully understand the purpose and the general idea, but not sure what the best workflow would be here, because I lack experience. Thank you!

[@saint85]

Is a flow hood really needed i want to get into tc but cost wise its a bit high looking for way to cheap it down just to get started and see if i want to keep at it was wondering if you or anyone can point me in the right direction thank you all

[@PlantCellTechnology]

Yes. It helps to maintain sterile conditions. But, if you are short on budget you can create a DIY hood for yourself using a box and plastic. (Watch the video on our channel to learn how to do that) But you will need to create a strict sterile environment for your cultures. and keep your hands clean too while working with cultures.

[@rajibmajumder843]

What we need add on rooting media?

[@PlantCellTechnology]

Are you asking what we need to add to rooting media? Or why do we need rooting media?

[@XiomaraS11]

Any suggestions on how to adjust the ph up and down without using sodium hydroxide or hydrochloride acid?

[@PlantCellTechnology]

Like are you seeking some other chemicals instead of NaOH and HCL?

[@XiomaraS11]

Yes, or could you proceed with the protocol without adjusting the pH?

[@XiomaraS11]

Can I use phenol red and bromothymol blue and how much of these?

[@PlantCellTechnology]

No, you definitely need to adjust the pH. @@XiomaraS11

[@PlantCellTechnology]

No these are staining chemicals. @@XiomaraS11

[@DuaneEseo-ul1bg]

I hope your equipments are available here in my country Philippines because I rather choose US brand compare to China.

[@PlantCellTechnology]

Yes, we ship all our equipment and chemicals globally.

[@scienceinfo9965]

Recently I have made agar media ,30 % of that media would not be solidify why????

[@PlantCellTechnology]

What concentration of agar did you use? Generally, you need to add 8 grams of agar to one liter of the media.

[@steamerpowered]

did you bring the batch up to full heat to melt the agar? its gotta go to around 90-95 deg F (ish) i had this happen the first few times i made media, didnt know it had to melt in, and had super random results, parts of the media were liquid, parts were super hard.

[@scienceinfo9965]

@@PlantCellTechnology we add 3gram agar and 4gram murashige

[@scienceinfo9965]

@@steamerpowered may be ,but I followed the same criteria which our senior follows

[@PlantCellTechnology]

If you are preparing one liter of the media. you need at least 8 grams of agar. So, that's why maybe your media is not solidifying well. @@scienceinfo9965

[@user-fw3zz5hh4i]

Can you please have video on how to add the explant into the media

[@user-fw3zz5hh4i]

Can you please have video on how to add the explant into the media

[@PlantCellTechnology]

We already have some videos on the channel. Alternatively, you may also see Francisco doing the procedure in many other culturing videos, such as "Introducing Philodendrons to tissue culture".

[@user-fw3zz5hh4i]

Can you please have video on how to add the explant into the media