Hey I did this a while back when I did my training. We followed an exact protocol by a commercial grower and none of us had any success, even though we followed the instructions to the last detail. With sterilization you only kill of bacteria, fungi, viruses and viroids on the surface of the explant. If there are fungi and bacteria growing within the plant you have little chance of a successfull in vitro culture. All you can do is transfer the infected explants again and again, sterilizing in between. Also, the smaller your explant, the higher your chance of success, since there is less material for fungi and bacteria to hold onto. Where I work we often use shoot meristems, sometimes only a fraction of a millimeter in size. This can prevent contamination. You can also try to be more and more agressive with your sterilisation, use longer and longer sterilisation times and stronger solutions. Hope this helps, keep on trying!
my first one was failure, but on the second try it kinda worked, but not on an orchid, the last time I did it, it was monstera Adansonii, it's still growing, it's even got a lil callus particles for now, the key point it to work in as clean environment as possible, contamination is the biggest issue in a TC, and also, try to play around with the scale of hormones and growth regulators, and find what works for you, that's the best way, you can't always act all by some research paper from fancy journal
@@quranrecitation6474 hey, I've not had any luck when it comes to the orchid tissue culture, cloning that particular specie of plant is quite complicated, you gotta experiment with media a little, maybe come up with your own mix or MS media, play around with hormones and regulators, also I'd highly recommend reading articles abt tissue culture and utilizing their experience
At that time I only used regular MS media with micro and macro elements, no hormones, but every plant has specific requirements, generally the exact protocol will be written in research papers and publications.
when i was making this video, i was fairly new so i had no idea about what proportions were required for phalenopsis orchid, so i just bought the one already optimized for work on orchid. I'll send u one, if i find correct proportions of micro and macro nutrients on the web
At 3:45 are you saying "and *don't* use the bleach solution that ..."? The mumbling speech gets in the way a bit, I had to replay a few other sections over and over too before I could determine what you said :(
did I really say that? to be totally honest, I can't remember a thing, I filmed that video almost 2 years ago, and yes, I HOPEFULLY meant distilled, rather than dehydrated, that just sounds too stupid, Thanks for mentioning (:
jars were heat sterilized before putting in the enclosure, abt enclosure, i bleach spraied it and than wiped whole surface with ethyl alcohol, btw i think i had 70% alcohol inside the enclosure, tho i'm not 100% certain.
So how can you take the gene that creates ultra violet colors and introduce that DNA into a plant so that it glows in the dark? Using at home plant tissue culture.
agrobacterium infiltration or gene gun, by agrobacterium, u use a plant pathogenic bacteria to introduce new dna (plasmid in this case) to a plant cells, by gene gun u literally shoot the genetic material into a plant tissue, than u tissue culture the targeted tissue and if successful, protein will be expressed and the plant cells will begin to be fluorescent under the UV light, in this case we'd use GFP - Green fluorescent protein "OR" RFP - Red fluorescent protein and many other options.
Super nice video! A few questions: 1. Which medium have you used? 2. Does boiling everything in the pressure cooker ruin something? 3. Did you use phyto hormones? I hope you continue to upload videos
Hi (: 1. I used Pre Mixed "Orchid Flask Medium 0753" on ebay, it's pretty simple medium containing Jelling agent and BA 2. I guess not! the only thing that is gonna ruin if u pressurecook* it , is live cells, they'll all die AND besides that, if u reautoclave your growth medium OR didn't adjust PH, it'll not solidify 3. i can't tell u that really, i literally didn't use anything beside that medium P.S: i'm beginner so a lot of things i'm doing may be inaccurate.
I havnt done this at home but i did it in Lab. First of all, you need to quarantine the explant before you take the shoot. Quatantine could take up to 3 months. Take care with regular fungiside and bactericide in semi steril environment. This will add successful tissue culture
@@agnediciuniene9861 I succeed on monstera species, you can check the newer vid as well, however orchid was lil rough, I still have to find a good multiplication mix for that, sorry for late reply
Thank you for sharing! I am amaze for such a young age and so passionate about your work. Please post the updates on tissue culture
Thanks, i will (:
Hey I did this a while back when I did my training. We followed an exact protocol by a commercial grower and none of us had any success, even though we followed the instructions to the last detail.
With sterilization you only kill of bacteria, fungi, viruses and viroids on the surface of the explant. If there are fungi and bacteria growing within the plant you have little chance of a successfull in vitro culture. All you can do is transfer the infected explants again and again, sterilizing in between.
Also, the smaller your explant, the higher your chance of success, since there is less material for fungi and bacteria to hold onto.
Where I work we often use shoot meristems, sometimes only a fraction of a millimeter in size. This can prevent contamination.
You can also try to be more and more agressive with your sterilisation, use longer and longer sterilisation times and stronger solutions.
Hope this helps, keep on trying!
my first one was failure, but on the second try it kinda worked, but not on an orchid, the last time I did it, it was monstera Adansonii, it's still growing, it's even got a lil callus particles for now, the key point it to work in as clean environment as possible, contamination is the biggest issue in a TC, and also, try to play around with the scale of hormones and growth regulators, and find what works for you, that's the best way, you can't always act all by some research paper from fancy journal
Hey can you help me with this tissue culture technique of any orchid? If yes then plz tell me your Instagram ID
@@quranrecitation6474 hey, I've not had any luck when it comes to the orchid tissue culture, cloning that particular specie of plant is quite complicated, you gotta experiment with media a little, maybe come up with your own mix or MS media, play around with hormones and regulators, also I'd highly recommend reading articles abt tissue culture and utilizing their experience
I congratulate you, although you are quite young, you have succeeded in a job that is quite difficult and requires knowledge. A big like from me.
Thanks (:
Plz make video on anthurium tissue culture
Thank you for upload by the way, it's always good to learn from the mistage. I wish I could learn this stuff when I was young as you.
That's a very practical video. Can you also please tell us the contents of the medium?
At that time I only used regular MS media with micro and macro elements, no hormones, but every plant has specific requirements, generally the exact protocol will be written in research papers and publications.
Your channel is great!
Thanks for sharing this info. Regards from chile 👍
glad you like it, thanks for the comment!
Hello can you give a components of tissue culture?
would love to know your media chemical composition
when i was making this video, i was fairly new so i had no idea about what proportions were required for phalenopsis orchid, so i just bought the one already optimized for work on orchid. I'll send u one, if i find correct proportions of micro and macro nutrients on the web
At 3:45 are you saying "and *don't* use the bleach solution that ..."? The mumbling speech gets in the way a bit, I had to replay a few other sections over and over too before I could determine what you said :(
By 'dehydrated water' you presumably mean distilled?
did I really say that? to be totally honest, I can't remember a thing, I filmed that video almost 2 years ago, and yes, I HOPEFULLY meant distilled, rather than dehydrated, that just sounds too stupid, Thanks for mentioning (:
Thank you very much ✌️😍
How are you sterilizing the jars. Also, I don’t see an IPA spray bottle in the enclosure. How are you sterilizing the enclosure after you seal it?
jars were heat sterilized before putting in the enclosure, abt enclosure, i bleach spraied it and than wiped whole surface with ethyl alcohol, btw i think i had 70% alcohol inside the enclosure, tho i'm not 100% certain.
I want to learn how to do that can you gave me the school
So how can you take the gene that creates ultra violet colors and introduce that DNA into a plant so that it glows in the dark? Using at home plant tissue culture.
agrobacterium infiltration or gene gun, by agrobacterium, u use a plant pathogenic bacteria to introduce new dna (plasmid in this case) to a plant cells, by gene gun u literally shoot the genetic material into a plant tissue, than u tissue culture the targeted tissue and if successful, protein will be expressed and the plant cells will begin to be fluorescent under the UV light, in this case we'd use GFP - Green fluorescent protein "OR" RFP - Red fluorescent protein and many other options.
Whats the name of the medium?
it's an MS medium optimized for orchid multiplication, every specie requires lil different optimization, all are murashage & Skoog though.
4:53 Your knife is too blunt.
Hello, your medium what are,the thinngs you put inside how Manny gram agar, water,enz
hey! i used a pre-mixed multiplication medium, here is the link if u like:
www.ebay.com/itm/292916657569
@@syntheogenesis01 Okay I will check it out
You see the rip in the glove right? I bet you didnt have a big enough fan also
oh ya, that's probably one of the reason all of the cultures got contaminated xDD + didn't have PPM that time in order to reduce contam
@@syntheogenesis01 Aye at least your at it, and that young.. You'll see many great things earlier than most.
Super nice video!
A few questions:
1. Which medium have you used?
2. Does boiling everything in the pressure cooker ruin something?
3. Did you use phyto hormones?
I hope you continue to upload videos
Hi (:
1. I used Pre Mixed "Orchid Flask Medium 0753" on ebay, it's pretty simple medium containing Jelling agent and BA
2. I guess not! the only thing that is gonna ruin if u pressurecook* it , is live cells, they'll all die AND besides that, if u reautoclave your growth medium OR didn't adjust PH, it'll not solidify
3. i can't tell u that really, i literally didn't use anything beside that medium
P.S: i'm beginner so a lot of things i'm doing may be inaccurate.
I havnt done this at home but i did it in Lab. First of all, you need to quarantine the explant before you take the shoot. Quatantine could take up to 3 months. Take care with regular fungiside and bactericide in semi steril environment. This will add successful tissue culture
Очень интересно 🥰
What plants did you succeed with? None of them look like orchids.
if you're saying abt thumbnail, they're not orchid, i just got it online.
@@syntheogenesis01 so, did you yourself have any success? What species was it?
@@agnediciuniene9861 I succeed on monstera species, you can check the newer vid as well, however orchid was lil rough, I still have to find a good multiplication mix for that, sorry for late reply
I m also a scientist .....doing job at tissue culture lab ...... contamination grow always how I removed it
some folks use ppm, it's quite effective, how do you deal with contamination?
👍👍
👍👍👍👍👍👍👍
👌👌
wow so inspiring
Thanks (:
You shouldn't cut a scalpal towards your hand like that. Cut away from you.
Not brand new tech, been used for 100 years
Yup, u're right, my bad.
kurang steril prosedurnya..tumbuh jamur
Tissue Culture was developed in 1895. Definitely not a new technology
If you compare the amount of time since the humans are developing agriculture versus the years between 1895 and today, its new tech..
You audio is not consistent, low and high. Put life to your voice.