Thanks for sharing! Can you talk more about the multiplication process? For example, if you start with one section of explant do you end up with one tissue cultured plant? I'm new to this but I though that the growth hormones result in multiple shoots from one explant which can then be rooted a second medium to produce many plants from one explant.
Hello, I watched video and couldn't find where you say what the ph should be for the media? I have read lots of protocols online and they dont tell me the ph. Can you tell me what it should be? And I am very curiuos to know if your anthurium worked out?
Hello there! Very informative. Thanks. I was wondering if its not necessary to cut the edges of the sterillized materiais, since I didnt see you doing that after the sterillization. As to my assumption, the edges are dead tissue, so I was wondering if they can still take advantage of the medium.
Hi, couldn't get which ends you are talking about. We took the nodes from where the leaves of the plants were grown (generally 2-3 cm of explants work), and then surface sterilize it and grew on nutrient-rich media.
@@PlantCellTechnology Hi! After the sterilization process, dont you have to cut the edges of the already exterillized explant? those exposed parts that got into direct contact with the bleach. they usually become pale/transparent
@@alvaromarra-e4i yeah, ideally you can do that. Or the most important part is removing bleach traces by thorough rinsing. This allows your plants to grow properly.
@@PlantCellTechnology I have tried both! Piper nigrum seeds had fungal contam after 50% bleach for 15 mins, twice! Nodes of P. longun had contam too. I think I might try your method of longer and less bleach. Have you tried suspension instead of agar after sterilization? I might try that too.
Great content! Could you kindly explain why you decided to add the hydrogen peroxide and cleaning vinegar? I have not found many sources that back the use of either with experimental data. Is the vinegar helping to remove bleach and H ₂O₂? Also, how can you be sure that they are sterile? Does adding these not increase the risk of contamination following bleach treatment? One more question: when you say 1% bleach, is this 1% of household chlorox (7.5% sodium hypochlorite)?
I have the same question regarding the bleach. Looking at the video it's per volume as he's filling the flask with 200ml water and 2ml bleach. I love the video's btw.
Edit: i found that this might be dangerous, do not attempt. I did find a secondary source which states that mixing hydrogen peroxide and vinegar produces an oxidising agent more potent than bleach. Is this correct?
Hydrogen peroxide and vinegar make a very strong oxidizer and a very effective sterilizing solution. The original formula is called "Vinoxide-HTC" HTC for home tissue culture. That formula is one part vinegar and four parts hydrogen peroxide. That solution is so strong. It can kill explants in less than 30 seconds. Over the years, I have been doing different experiments and rinsing the explants for 5 minutes with hydrogen peroxide, and then adding the vinegar has given me the best results.
Love the work. Thanks for looking at the request.
Any time!
Could you please explain by what you mean with splitting some growing points?Like do you cut them open ?
Did you transfer the explants right out of the peroxide/vinegar solution?
Yes, no need to rinse!
Is it possible to use leaves as an explant? Thanks.
For Anthurium will be difficult as you have to get callus first then regenerate plants from there. That is harder than it sound.
Thanks for sharing! Can you talk more about the multiplication process? For example, if you start with one section of explant do you end up with one tissue cultured plant? I'm new to this but I though that the growth hormones result in multiple shoots from one explant which can then be rooted a second medium to produce many plants from one explant.
Hello, I watched video and couldn't find where you say what the ph should be for the media? I have read lots of protocols online and they dont tell me the ph. Can you tell me what it should be? And I am very curiuos to know if your anthurium worked out?
When you put the explant in hydrogen peroxide was it diluted by water or straight from the bottle?
Hydrogen peroxide was only used to rinse the explants after sterilizing them in 1% bleach.
Hello there!
Very informative.
Thanks.
I was wondering if its not necessary to cut the edges of the sterillized materiais, since I didnt see you doing that after the sterillization.
As to my assumption, the edges are dead tissue, so I was wondering if they can still take advantage of the medium.
Hi, couldn't get which ends you are talking about. We took the nodes from where the leaves of the plants were grown (generally 2-3 cm of explants work), and then surface sterilize it and grew on nutrient-rich media.
@@PlantCellTechnology Hi! After the sterilization process, dont you have to cut the edges of the already exterillized explant? those exposed parts that got into direct contact with the bleach. they usually become pale/transparent
@@alvaromarra-e4i yeah, ideally you can do that. Or the most important part is removing bleach traces by thorough rinsing. This allows your plants to grow properly.
did this protocol end up working
Oh yes! It worked in our lab.
Can you post the next videos of this ?
Thank you very much for sharing
Can you do a video on the different types of PGRs sometime? Thanks.
Yes, I'll do a video about PGRs soon.
What did you add in 2.22? What is a brown bottle?
3% hydrogen peroxide from the pharmacy.
@@PlantCellTechnology Thank you.
Please do a videos on multiplication too..
Thanks you
Great idea! - Working on it! Should be coming soon!
@@PlantCellTechnology Waiting 👍👍
I keep getting fungal issues with Piper species. Can you try black pepper? (Piper nigrum)
Are you working with seeds or plants?
@@PlantCellTechnology I have tried both! Piper nigrum seeds had fungal contam after 50% bleach for 15 mins, twice! Nodes of P. longun had contam too. I think I might try your method of longer and less bleach. Have you tried suspension instead of agar after sterilization? I might try that too.
Great content!
Could you kindly explain why you decided to add the hydrogen peroxide and cleaning vinegar? I have not found many sources that back the use of either with experimental data. Is the vinegar helping to remove bleach and H ₂O₂? Also, how can you be sure that they are sterile? Does adding these not increase the risk of contamination following bleach treatment?
One more question: when you say 1% bleach, is this 1% of household chlorox (7.5% sodium hypochlorite)?
I have the same question regarding the bleach. Looking at the video it's per volume as he's filling the flask with 200ml water and 2ml bleach. I love the video's btw.
If my calculation is correct his concentration of bleach is 0,074% if his bleach from the bottle is 7,5%
Edit: i found that this might be dangerous, do not attempt.
I did find a secondary source which states that mixing hydrogen peroxide and vinegar produces an oxidising agent more potent than bleach. Is this correct?
Hydrogen peroxide and vinegar make a very strong oxidizer and a very effective sterilizing solution. The original formula is called "Vinoxide-HTC" HTC for home tissue culture. That formula is one part vinegar and four parts hydrogen peroxide. That solution is so strong. It can kill explants in less than 30 seconds. Over the years, I have been doing different experiments and rinsing the explants for 5 minutes with hydrogen peroxide, and then adding the vinegar has given me the best results.
Thanks, and that's correct.