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@@greenhut1642 addition of activated charcoal would help

@@afiqfauzan2941 yes, all kinds of ginger family also possible…but, you may need some adjustments on the sterilization protocol and media

@@Yoya974 2 weeks…as this added BAP

Thank you. Where can I buy reliable seeds, because after my tests, my seeds do not germinate? Please !

@@Yoya974 you may just go nursery and ask, or neighborhood…do you remove seed coats?

Yes! I removed it. The samples were first inoculated on MS medium without charcoal with BAP, without result. Then on MS with charcoal and BAP, already 4 weeks ago.

@@Yoya974 try to put it another month or two…sometimes it takes longer time.

@@JonesyMytube then don’t watch ☺️

@@JonesyMytube mute it then ☺️, fingers are yours

@@Mukesh12-ob9nb No, it depends on your needs

@@himanshigaonkar6300 Hi, dear, you just visit the website Plant Cell Tech will do☺️ Please remember that, 1 code only for 1 use, if you’re going to buy it, make sure you buy in bulks to save more ☺️

@@spineless5990 you have to test it out… generally, all PGRs are the same to me regardless of the brands

@@kralice462 you can have any X (times) you want, then prepare the volume accordingly… As long as you calculate using dilution formula, M1V1=M2V2 correctly, it won’t be an issue ☺️

Can you state the time in the comment?

@@Plantastic12345 27:24 media on petri which media u using? just ms media ?

@@jedz86 yes, it’s MS basal media. At this stage, we usually don’t add any hormones as there might be contamination. Moreover, the growth is super fast once it’s germinated without contamination.

Yes, it does induce multiple shoots with minimal amount of BAP. You can multiply after that ☺️

@@Plantastic12345 thanks for the answer, maybe you can do a follow up for other cereals

@@Vgb_ali_official if and only if I have the access to lab and also the permission of making the process “transparent” in future 😅

@@Plantastic12345 okay thx

I think you could refer to any journals on banana you want, then purchase ready MS media and add the hormone respectively

Yes, it is, but it does produces desired results for research purposes

English please

@@Plantastic12345 0,022 gr hormon/ 1L water ?

@@Человекновоговремени nope, you may refer to 0:15… It’s mentioned that. 0.022g of TDZ is used to prepare 1.0 mM of TDZ at 100 mL If you want to prepare for 1L, you just x10 of the amount needed. But I’m not recommending to do so, as stock solution is always prepared in small volume but high concentration.😇

@@Plantastic12345 So 100ml is the mother solution of the substance? Then how much of the mother liquor should I take for 1 liter of nutrient medium?

@@Plantastic12345 So 100ml is the mother solution of the substance? Then how much of the mother liquor should I take for 1 liter of nutrient medium?

🥹 This is a very good example: Never put anything near to the weighing boat! 😂

I had finished uni study and no company would ok with videos taking

You need some moisture to make sure it’s fertilized between individuals… I didn’t proceed with further stage as this is the stage that client wants as they mentioned that induction of thallus phase in nature is very slow

If the seed pods are unopened or without any cracks, you may perform as the same shown in the video.

這些都需要加一些碳粉和antioxidant在初期，之後就可以依據一些文獻

I am from Malaysia. Thank you for being so supportive. I am trying to make more content in the future.

Endogenous bacteria and fungus are possible in any explants… If you had diagnosed your plants already infected with such, then you need to use meristem tissue culture…

Use NaOH or KOH then

@@Plantastic12345 thank you sir

Filter pore diameter is depending on the chemicals itself… You need to search what’s the pore diameter required for you to do the filteration…commonly 0.22 or 0.45 micro meter filters will be used depending on the chemicals… Hormone labeled with Ca/F is either they can be autoclaved or filtered… For more effective usage, I usually do filteration if there’s a choice between autoclave and filteration…as most paper discussed that high temperature of autoclave may reduce the efficiency of the hormones

@@Plantastic12345 yes i understand this. thanks for your reply. As I'm using IBA,IAA,K-IBA,TDZ,and KIN i think 0.2um is what i need. please let me know if I'm wrong. however my question was about the diameter of the filter , such as 33mm vs 17mm etc. depending on volume , but for making stock solutions which do you recommend?

You need to check what solvent to be used…and yes, most of the common hormones for tissue culture can be used as solvent… Just google the solvents for the particular hormone then you will get the answers

@@Plantastic12345 yes thank you, at the moment getting: k iba then BA, IAA, TDZ, KIN, NA all can use KOH

有，是可行的… 基本上它都是跟我們用不同的鹽炮製的是一樣的成分… 準確度跟時間方面來說，它是很方便！

這個是調制好的，直接衝泡就可以了… 目前只有Murashige & Skoog 培養基哦！

他的成分都是跟用個別的鹽調成的培養基都是一樣的…只是這個適合小型的實驗室

Plant and tissue culture grow differently, as plant growth regulators stimulated the growth of tissue culture plants

1. PPM is strictly depends on the conditions of the explants (spores in this case). You may use 5, 10, 15, and 20 mL per liter in the gelled media as I’m not sure if the growing condition of the plant bearing spore is “clean” or not. 2. More than an hour. 3. Yes, same. 4. Quick dip in 70% ethanol before Clorox. The concentration of Clorox is strongly dependent on the amount of the spores per tea bag and the penetration of the tea bag. 5. All sterilization process in my videos are: Tap water> 70% ethanol> Clorox> rinsing with sterile water. The reason I don’t put concentration used and duration is because everything has to be adjusted based on the explants we used and the status of the explants.

@@Plantastic12345 Thank you! I'll try it again and let you know results in 1-2 months. Ive already noticed few of the spores from previous experiment is green now :) thanks

Hi, Daniel. You may use it for all type of plants. As every hormones need to be optimized as well as types of explants and concentrations need to be tested… For exact or approximate range to be used, you need to search some journals

@@Plantastic12345 Thanks a lot for your reply. Wanted to use it for roses and anthuriums. I will appreciate if you have any recommendations.

@@danieljheelan5256 never work with those plants before, but I’m pretty it’s very easy to get reference from online…if you need consultation, you may let me know, thanks

@@Plantastic12345 Thanks a lot.

Depends on the hormone you used…you may refer to some journal… It took about 2 months from my past experience based on this video

Which hormone you used for callus induction....I am using 2,4-D only for rhizome of curcuma

@@ahlamabdulaziz6513 I used 20 micro molar BAP

​@@Plantastic12345sir please tell me why we use hormones

@@ZahidButt-f8g Speedup growth and development and thus reduce waiting period

For the solvent, it’s fine so far from what I had been using… It won’t bring much effect to the media since we used only very little amount of the stock solution… That’s why, it’s better to prepare very concentrated hormone stock solutions than a less concentrated one…

@@Plantastic12345 thank you for the detailed reply. What about that affect on the plant tissue? I guess when it is mixed with media it will be diluted enough where it doesnt matter?

@@NagashiChidorii we never know the pH of the media after sterilization…therefore, we usually fix the range of the pH from 5.7 to 5.8 before sterilization… For such method, I fix the pH at 5.7 in case the very little hormone increases the pH slightly…

@@Plantastic12345 after sterilization, i read that PH will drop due to acidification mechanisms from the heat. I tested this last night and it seems to be true. I PH my media with meta-toplin to 6.2. After sterilization i tested Ph again and it dropped to 5.3 or 5.4. So try checking that. I just ordered NaOh 0.1 N and TDZ thanks to your video. Thank you! Now I wonder about 0.5 N… How do I measure 0.5 uM TDZ in my media?

@@NagashiChidorii actually there’s no need to measure pH in stock preparation and the media after sterilization

嗨，您好。 蕨類植物的我也是用MS培養基哦…… 我建議您調稀一半的試一試。

@@Plantastic12345 請問鹿角蕨你有，用組織栽培過嗎？

@@游泓清 我试过用它的孢子，一样的…你可以看fern tissue culture视频

用其他部位能夠培養成功嗎？

如果用其他部位可以的話，可以有條件的請你幫忙嗎？

Wipe the surface of the pods using ethanol then keep them in a container inside a fridge for cold storage… Alternatively, you can keep it at shaded dry area… However, both are not recommended as we need to culture as fresh as possible when we harvested it

@@Plantastic12345 for long it can be kept with the methods you mentioned without loosing its viability?

@@amintaleghani2110 I only tried to keep for a week…did not try to keep longer than that…

Diluted Clorox at 1-10%

@@Plantastic12345 Then rinse with sterilized water?

@@AquaMayne yes