How to Make Plant Tissue Culture Media for Beginners: Step-by-Step Tutorial
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- čas přidán 19. 01. 2022
- In this video, our lab director Francisco will guide you through the process of making plant tissue culture media from scratch. Tissue culture media is an essential component of plant tissue culture, providing the necessary nutrients and growth factors for the successful propagation and maintenance of plant cells and tissues in vitro. Get 10%OFF the tissue culture products you need using my code: www.plantcelltechnology.com/?...
Whether you are a student or a researcher new to tissue culture, this tutorial will help you understand the ingredients, preparation, and sterilization techniques required for making high-quality tissue culture media. Francisco will demonstrate the step-by-step process of making a basic Murashige and Skoog (MS) medium, including the proper measurement and mixing of ingredients, pH adjustment, and autoclaving. You will also learn about the common mistakes to avoid and the tips for troubleshooting any issues you might encounter. By the end of this video, you will have the confidence and knowledge to prepare your own tissue culture media and start growing healthy plant cultures in the lab. So, let's get started!
Don't forget to subscribe to our channel for hundreds of free educational plant tissue culture videos and be sure to mark your calendar for our upcoming Houseplants Tissue Culture Master Class with Francisco in Washington DC. This is a unique opportunity to learn from the experts and take your plant tissue culture skills to the next level. Join us and discover the amazing world of plant tissue culture today! plantcelltechnology.com/master-classes/
We are Plant Cell Technology, a leading plant tissue culture industry organization since 1995. We develop, produce, and market innovative solutions for global plant tissue culture operations and hobbyists. Our well-researched, certified, and trusted products keep your cultures healthy and contaminate-free.
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This open source information is so important. Thank you so much for sharing. Im heading over to the website to support and pick up supplies.
Thanks for your kind feedback and support. Let us know if you need any other help in your tissue culture journey.
This is gonna open up some GREAT opportunity to finally desaturate the “rare” plant market, and make rare plants accessible to more people!
It will be accessible but not cheaper to access because of the cost involved. So, rare will still be rare.
Are you interested in tissue culture to grow rare plants?
@@PlantCellTechnology yes, my goal is to create and open a ship that sells “rare” plants at affordable prices, and I think TC could be the ultimate way to accomplish that.
@@isaacspeyer1129 Definitely! The market of rare plants is expanding and the demand is still needed to be fulfilled at that scale. So, it's a great idea for your business. Good luck!
Let us know if you need our help in your journey in anyways!
Hi first of all than you very much for taking the time to make this informative video! For the step where the Hellman gum is added to the left over solution and placed into the plastic container. Is that solution put into the biocoupler? Or is that kept aside for later? I am not sure I understood when that solution is needed for the plant extract. Thank you for your time!
After 12 hours on the internet researching this subject... your the first to actually show every detail. 👏 👏
Thanks, Dylan. We also have blogs on the subject in case you are interested to learn about some topics. Here's the link: www.plantcelltechnology.com/blog/
@@PlantCellTechnology dude for real. Your the top Tissue culture guy on CZcams at the moment. Easily the most informative, best delivered, and then respond with a whole slew of more info in blog format. You rock dude. 👏 I'm not subscribed to many people. Deff with your channel tho.
@@Dylan-lt9eo Hey,
Thanks for your appreciation, Dylan!
But, the blogs on the website are written by our scientific writer, Anjali Singh.
She also helps other culturists with their tissue culture questions, which you guys send over in the comments or message us on our social media.
You can give her kudos for her blogs or ask her questions by reaching out at anjali@plantcelltechnology.com.
When sterilizing in a pressure cooker an empty jar should have a few drops of distilled water inside. The steam is essential to help denature contaminates as well as push out air. Some microorganism spores can survive temperatures far above 121c when in a dry atmosphere, The air also causes uneven temperatures.
In a true autoclave they have much better sealing and the steam is generated externally and added at the top, with a vent at the bottom because water vapor is less dense than O2 and N2. Then the autoclave will start with a series of pressure and venting cycles to purge air from all cavities and fill them with steam. (Some models may start with a vacuum depending on the intended material to sterilize.) This purging of air is also a way to ensure the temperature in all corners, if there is 100% H2O vapor and 15psi then any cavity below 120c will condense and draw in more hot steam. However if there are gases like N2 or O2 they remain gas at lower temperatures and will prevent much of this condensing heat transfer, and gases are poor conductors so reaching an even temperature may take a much longer time.
Thanks for your insight!
Thanks so much for this!
Glad you liked it!
Does the biocoupler lids fit a regular mouth canning jar or a wide mouth canning jar?
When you put a plant into tissue culture for the first time, in order to get the plant initiated for the first time in tissue culture, do you use half (or even a 1/4) the amount of plant growth regulators that are called for in the protocol you are using? Thank you!
Do you use both the rooting hormone and the branching compound together when making the tissue culture medium?
Awesome video!
Happy to know that you found the video useful.
Can you use clonex to put in there for your root hormone? It would be nice to have a list and of how much in the description or an easier "ghetto" version of chemicals you need.
I was thinking the same thing. I was considering mixing cloning solution in with agar...
Great video man
Thank you so much!! Stay tuned to our channel to watch more videos like this.
How can I calibrate my new pH meter? Could you please teach me how to prepare the NaOH and HCL solution required for calibration? İ
I very much admire your instruction. So ordered for your basic kit, but it did not come with your plant regulators. Should I order those separately or will it still work without them?
Yes, the kit doesn't come with hormones. You need to purchase them separately from our website based on the protocol you are following for your plants.
Hey buddy big fan !
Question, i am a medical cannabis cultivator and growing for 25plus years mainly hydroponics but in last 4-5 years focused on regenerative .
Growing hydroponically, i grew in more sterile ways . With organics i use diversity of microbes bacteria and fungi . I have been learning TC for a short time and started to wonder if my best option is to go back to hydroponics instead of organics when comes to adding TC to my methods of cultivation. Would it be easier transition when harden off the TC explants and when decide to store and propagate phenotypes i that are ones i want to keep .
I want to be able to mother without using the space id need to cultivate mothers . Also to preserve and store plants i choose. What do u suggest is my best cultivation method?
Oh btw i recently moved to apt from a house with big gardens .indoor and out .
is it possible to use normal LED light
I ordered the starter kit from ur website. Does that include the cytokinin and auxin , also im interested in a consultation do u do those over zoom? Thanks
Hey Jason,
It's so amazing that you bought the starter kit. The kit contains everything but plant growth hormones.
So, you have to purchase the cytokinin and auxin separately.
And, about consultation, yes we do it over zoom.
If you are trying to do tissue culture on a Christmas cactus, how do you lay the leaf? Do we stick the bottom of the leaf into the medium ,are can we lay sections of the leaf flat on the medium?
I haven't done TC with Christmas cactus, but I would stick the bottom of the leaf section on the media.
bro please tell me how to use albert solution for tissue culture and how to use natural things for hormones
Saludos Francisco. En donde estan ustedes. Quiero comenzar un proyecto para suplir mi finca y cuando este mas diestro algunos otros agricultores locales. Me quiero concentrar en tuberculos dado que mis colegas se quejan de escasez de semilla y de bananos. Seguro hare algunas ornamentales para diversificar pero inicialmente malanga, yautia y bananas. Estoy en Puerto Rico. Gracias por tus videos.
Hello
1. Can you help me to use pvp antioxidant in media (how much use it in M.S at intiation stage)
My plant is Opuntia cactus.
2. How is time with concentration to soaking explant in ascorbic and citric acid
3. Can I cutting my Cladodes before soaking or just cut it when I plant it in media .. thank you so much
Hi there,
We see you have some specific tissue culture questions regarding a specific plant of yours. We'd recommend booking a consultation call with our lab director who can help you answer all your questions instantly. Check out more details here: www.plantcelltechnology.com/plant-tissue-culture-consultations/
Hi. can you give me the link where you buy your deli containers? I always though plastic would melt or warp in the pressure cooker.
I buy it from my local store. You need to ask them to give you a container made of autoclavable plastic.
So the Plant Growth regulator is Cytokinin and Auxin?
Yes, these two hormones are mainly used for the shoot and root regeneration in tissue culture. But, the concentration varies based on the plant species you are working with.
Thanks for the info Francisco. Listen, what is a good media formulation for Cannabis?
For cannabis tissue culture, you can book a consultation call with us.
www.plantcelltechnology.com/pct-consultation/
@@PlantCellTechnology oh come on dude! I'm Colombian. I can't afford that! Give me a hand here please!!
@@randomsht196 Exact media formulation can only be provided by the culturists in the field. You can try approaching cannabis cultivators for your help.
I have a question. Is it safe to heat the ingredients , including plant hormones , in my microwave that I use to heat food up. I have heard that some on the plant hormones are so dangerous that you shouldn't touch them. I've been wanting to try tc so this would be my first time. I'm not a perfessional just someone interested in trying this out and seeing where it goes.
Any specific plant hormone that you can name. You can work with plant hormones easily with certain precautions as mentioned in the safety datasheet of products. Also, hormones are sterilized after adding them to the media, and for media sterilization, the autoclave is the most preferred technique. So, if you are starting out, try investing in a pressure cooker for tissue culture purposes.
Do you know how much ml/l of IBA for rooting should be used because i want to make cuttings from plants and i allready have IBA from you and do not want to buy anything else?Thank you.
The concentration of hormones differs based on the plant species you are working on. So, try to follow a recipe for the plant you desire to grow. In tissue culture, there's no one-size-fits-all concept. what plant are you growing?
Can you put a list of all ingredients and amounts needed in the the description
Oh sure! We'll do it soon.
Can you sell a pre mixed media. I just want to sterilize my cutting and drop it in?
That's a great idea! Though, currently, we aren't selling them. Maybe soon in near future!
awesome video! Is BAP dangerous to keep in your house? I don't have pets or kids or anything, but should a person wear a mask and gloves when mixing media containing BAP?
No BAP is not dangerous and safe while used in TC experiments.
Thank you!
where can I buy the hormones from, i cant see them on the plant cell technology site?
which hormones do you need?
@@PlantCellTechnology I am wanting both Auxins & Cytokinin, along with PPM & MS Media. Can you ship it "Door to Door"?
@@vinodthadhani5055 You can ask these queries at info@plantcelltechnology.com. You will also get the answer to your requirements instantly.
Can we use small amount of fungicide and antibiotics to disinfect the ex plants ?
You can use them but they might have certain negative effects on plant growth. Why don't you try PPM? It doesn't have any side effects and it can be autoclavable with media, unlike antibiotics.
@@PlantCellTechnology Thank you. Will try for sure
I'd love to get the kit, do you send to Colombia?
Yeah, sure!
I want to buy all the liquid that makes media,
Will adding the geling agent not affect the ph again?
Hey Dane,
Very nice question!
Since agar has no significant effect on the pH of the culture medium and doing this keeps your pH meter clean, it's advisable to measure pH before adding the agar.
However, there're certain labs that measure pH after adding agar.
To avoid your confusion by yourself, for one time you can measure pH both, before and after adding agar and see it by yourself!
I hope I've answered your questions!
Let me know if you have any other questions.
👍👍👍
Make a video on media preparation for Anubias pant
Sure! Until then, you can learn about it in our blog: www.plantcelltechnology.com/blog/how-to-tissue-culture-anubias/
6ba and naa powder i have, how much quantity should i take both
For what do you want to use the hormones.
@@PlantCellTechnology for tuberous plants like caladium,
Better to search for a working protocol for the plant and find details on every stage of the tissue culture process of the plant. @@greenhut1642
Hello sir im happy to come across your vedio,were can I order the media for potato tissue culture
Hi,
You can purchase basic tissue culture requirements from the PCT store: www.plantcelltechnology.com/pct-store/
But to formulate the media for potato species you need to find a specific potato tissue culture protocol.
do you sterilize the media that goes into the deli containers? if so how do you go about it.
also. what is considered heat safe hormone? i dont see a diffrence between any of them.
@@bcerrati we pour the media into containers and then put them into an autoclave for sterilization.
Heat safe or heat-stable hormone is the hormone that can be sterilized with the media., that is autoclavable.
And, heat-labile hormones are the ones that can not be autoclaved with the media and need to be added after the media is sterilized, using the syringe filter technique.
Here's a list of hormones that are heat labile and heat stable: www.sigmaaldrich.com/IN/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/plant-tissue-culture/growth-regulators
Do u have videos for making media for carnivorous plants
Yes! We have many available on our channel. Do check them out.
How should I do things to buy?
You can purchase all your tissue culture requirements from our website: plantcelltechnology.com/collections/all
PPM is hard to find here in India. What is the alternate that I can use ?
You can order PPM from plantcelltechnology.com or can contact our Indian distributor. Below is the address and contact number.
Jyothi Shikhar Tower, Professor Joginder Singh Marg, Kirti Shikar, Janakpuri District Center, Janakpuri, New Delhi, Delhi 110058, India
91-9810029475
biogenuix.com
An alternative is an antibiotic use. But, it has several limitations.
Thank you so much! Will contact your distributor in India.
@@TheOrganicSpade Great!! Let me know if there's anything else that you need help with. Happy culturing!!
How much would you use for media vanilla
What component of the media are you talking about?
Plant growth regulators and I also can't find he video to make sodium hydroxide for the PGR.
@@nikkilynick9606 Here's the video on NaOH: www.youtube.com/@PlantCellTechnology/search?query=naoh
You will need to search the papers on Google to discover the type of hormone required for the tissue culture of Vanilla plants as we do not work with them in our lab.
Sir, can we do tissue culture of dragon fruits??
Of course, you can do it! There're multiple studies available that report the successful propagation of dragon fruits using tissue culture.
@@PlantCellTechnology thanks sir 🌹
@@MuhammadFaisal-rk8cc Let us know if you have any other queries or questions. Happy Culturing!
@@PlantCellTechnology ,,🌹❤️🌹
Hello sir,
I have tried orchid callus induction experiment, all the processes, sterilization and other requirements are good enough for the cultures, but I can not see any callus induction in my Culture tubes ....
Could you please help me with this ....
Thank you 🙏
What procedure are you following?
@@PlantCellTechnology First of all, thank you for your consideration....
I'm following some research paper, which suggests using auxin cytokinin ratio ( 1:1, 2:1 , 1:2) .... And the hormones I'm using are IAA, BAP, kinetin, NAA, 2,4-D ....
Please suggest me something which could help me ...
🙏
@@kalpanarautela320 You can try to talk directly regarding this problem to our scientist. Because you can get a better solution that way.
You can book a call here: www.plantcelltechnology.com/pct-consultation/
Also, I can see that you are using different ratios of hormones and different types of auxin and cytokinin. So, did none of the combinations of these work for you?
@@kalpanarautela320 In that paper, you will find which ratio and type of hormone worked best for that particular lab. You can try that one. And, also make sure the plant species you are working with are the same as mentioned in the paper.
Because, if the species is different, the protocol might not work the same for you as it worked for that lab.
@@PlantCellTechnology okay ...
Thank you so muchh for your consideration... 🙏
Thanks for the video
Can you help me with the formulation of media for figs plz?
Happy to help! Currently, we aren't growing figs in our lab, however, you can some resources available on the internet. And, soo we might create a blog on the topic as well. So, we will definitely inform you as soon as it's live.
@@PlantCellTechnology thanks
But in theory do i have to grow TC figs in different medias like should i use media for multiplication first then cut the shoots and then use rooting media and then transfer them for hardening?
Sorry i am new to this so it might be a silly question
@@ayyazali4037 Theoretically, you should first use normal media without hormones to establish your cultures, then multiplication media containing shooting hormone for shoot development, and then rooting media containing auxin (the rooting hormone) for root development.
@@PlantCellTechnology great…thanks
I spoke to someone in my area to make me a media and he asked me to find a protocol for figs so can you explain what that means and can you guide me about the protocol for figs plz?
@@ayyazali4037 Protocol is the procedure or steps you need to follow to grow plants in tissue culture. It mentions what chemicals you need and in what concentration and how to proceed with the tissue culture process.
You can find the protocols in research papers available over the internet.
Here's one example that I found:
www.ncbi.nlm.nih.gov/pmc/articles/PMC6072732/
Like this many others will be there. You can have a look at them.
I hope it helps.
what is this biocoupler for?
The Bicoupler is used to multiply the established in vitro or tissue culture plants.
Is there a list anywhere with all the things you used in the video that you guys sell on your website? I need all of this stuff asap
On our website, you will get everything to prepare media, including MS media, agar or gellan gum, and PPM.
Additionally, you can also purchase vented glass culture vessels and Biocoupler TM.
Here's the link to PCT Store: www.plantcelltechnology.com/pct-store/
I'm not sure if i missed it in this video but when is the agar added?
Gellan gum is added at the last of the video, which is also a gelling agent like agar.
@@PlantCellTechnology does gellan gum needs to be heated or does that applies only to agar?
@@TheJavo123 You will have to heat it. The gum is also insoluble in cold water.
can you tell me how much agar I would add for 1 litre of medium
6-10 grams
Will this work for hemp plants?
The steps shown in the video are basic steps to prepare a tissue culture media. However, for a specific plant, you need to experiment with the type and concentration of plant growth regulators to see which suits best for your plants.
You can also look for research papers in which protocols for tissue culturing cannabis plant is given.
@@PlantCellTechnology Thank you great idea!
@@Degen_Martyr
If you are a beginner in cannabis tissue culture, then you can try our consultation services to get instant solutions for your problems. You can use this link to book the call: www.plantcelltechnology.com/pct-consultation/
I guess where I find myself confused is media for shoots and then for the roots
awesome, i think they call it a turkey baster
cuanto cuesta el kit?
Here are the details: www.plantcelltechnology.com/starter-kits/
Can you share info on how to troubleshoot tissue cultures? I have tried culturing begonia explants, they seem sterile but are not growing after over a month. I had 2 explants produce a fairly large leaf, but then after i transplanted them, again, no growth. I've tried various ratios/concentrations of growth hormones but it didn't seem to make a difference. I also tried 50 and 100% concentration M&S media. Any tips?
We published a blog on the tissue culture of Begonias. You might find that useful. Here's the link: www.plantcelltechnology.com/blog/tissue-culture-propagation-of-begonias-/
@@PlantCellTechnology Yes, I actually followed exactly your guide for my "baseline" recipe/protocol, tried with several types of explants even. I'm not sure why it didn't work - do you have any examples of what it looks like when it's working? How quickly should it grow and stuff?
@@MagnusNyborgMadsen Unfortunately not! We don't have any ongoing experiments on the plant.
What is the advantage and disadvantages using the high cost and low cost medium?
when you say high-cost and low-cost medium, why medium do you refer to? Can you be a bit clear on your question? It might help us to better understand that.
Thanks!
@@PlantCellTechnology I mean, i am can ducting a research paper, my title is Low cost medium for rapid in vitro propagation of banana Var. Cardaba(Musa acuminata × balbisiana) so I saw that in tissue culture media the price is high, but I thought if I could reduce the standard compositions, do you have any thoughts to lower the tissue culture media.
@@ynercabigquez9531 You can use cheaper components in the media that impart the same value to the plants. However, in tissue culture effectivity is a major concern. So, if you are trying to reduce the tissue culture media cost, try to replace it with the components that are effective as well, and that do not impact negatively the growth and yield of the cultures.
@@PlantCellTechnology I see, by the way, what is the impact or help the tissue culture on crop protection major?
@@ynercabigquez9531 Tissue culture helps you to get rid of diseases and increase plants' overall yield.
If you perform the process in a complete aseptic condition, you will be surprised by the healthy growth of plants.
Do you guys sell the Heat Stable Plant Growth Regulators?
No!
Can you name all the material needed to start this Tissue culture mediums,and the other question is can I use this media on potato tissue culture,third question were can I buy these raw materials in South Africa
1. Check out these articles to learn all about starting in tissue culture: www.plantcelltechnology.com/search.php?search_query=beginner§ion=content
2. You can definitely use basic materials such as MS media, agar, PPM, and hormones for potatoes. however, you need a specific tissue culture protocol to introduce the specific plant species into tissue culture.
We have certain blogs on it, check them out: www.plantcelltechnology.com/search.php?search_query=potato§ion=content
3. You can either buy from our website www.plantcelltechnology.com or we also have a distributor in South Africa.
Here're the details:
Aquarium Corporation of South Africa
Main Office
Gauteng, South Africa
27123291714
moolis@aquaticplants.co.za
Were can I buy your products in South Africa 💖💞
@@agrippanyagwaya2308 You can either buy from our website www.plantcelltechnology.com
or we also have a distributor in South Africa.
Here're the details:
Aquarium Corporation of South Africa
Main Office
Gauteng, South Africa
27123291714
moolis@aquaticplants.co.
Thank you for your support
@@agrippanyagwaya2308 Of course! Happy culturing!
Can these measurements be used for cannabis?
No! You will have to formulate it according to the plant.
If you are a beginner in cannabis tissue culture, then you can try our consultation services to get instant solutions for your problems. You can use this link to book the call: www.plantcelltechnology.com/pct-consultation/
Please list the ingredients.
Hey! Sure we'll do it soon. You can refer to this blog as well for media preparation.
www.plantcelltechnology.com/blog/for-beginners-requirement-and-preparation-of-growth-media/
Thank you for your response. So If I buy the starter kit with the jars, PPM, Agar, and MS media I can make the media. Will this media be good for philodendrons, Anthurium, Alocasia and Hoya? What about BAP and the other chemical you used. That’s why I’m a little confused. Thank you.
@@jwcollect8724 For establishing the cultures starter kit will be great but for the multiplication, you'll need plant growth hormones, such as BAP and others.
Just to give you an idea, here's the recipe for Philodendrons that work in our lab:
For multiplication:
Full MS media
25 g of sucrose
3-5 ml/l of BAP
0.3-0.5 ml/l of NAA
1 ml/l of PPM
6 g of Agar or 3 g of Gellan gum
For rooting:
Full MS media
25 g of sucrose
3-5 ml/l of NAA
0.3-0.5 ml/l of BAP
1 ml/l of PPM
6 g of Agar or 3 g of Gellan gum
@@PlantCellTechnology Hi, Are you saying that after roots appear you move the plants to the other formula to grow? Is there a compromise formula so you don’t have to switch? Do you sell BAP and NAA? Thank you for all the videos, I’m learning a lot. Thanks.
Hi, I think I got it. You make the basic media formula, then add Auxins, Cytokinins and Gibberellenes depending on rooting, multiplying or growing.
Not to give away secrets can you steer me to where I could get more info on adjusting the formula. Are there different formulas for different plants like Anthurium, Hoya, Alocasia and Philodendron? Thank you very much for all your help, videos and info. Regards.
why you all cross armed at 21 minutes?
Couldn't get your question. Can you please reframe it?
Very informative thank you very much
Thank you so much for your comment, Said!