Who is our resident Tissue culture master?

[Moe.Red]

Subscribed. Another thread I need to start at the beginning.

 

[Dr.Mantis]

I'm really happy to hear you say that,,,im all about helping individuals make some progress ,,,when it comes to personal people doing personal research i can't support you enough,,

DKW displays improved callogenesis compared with MS basal salts in some cultivars

What have you read so far?

Thank you!

It’s always something I’ve wanted to get into. Now that I see others have done it, I know it’s possible.

The DKW idea came from this paper: https://cdnsciencepub.com/doi/10.1139/cjb-2020-0179

This actually worked out well for me, because I didn’t really want to buy MS media and I already had the DKW raw ingredient and vitamins for gamborg vitamins.

I read another paper somewhere that mentioned silicate (6um I think?) improved shoot rooting. I’m not sure where to start on the hormone side. I have tdz, gib, naa, kin, and bap. I was thinking tdz or tdz + gaa for shoot generation. Then perhaps naa + silicate for the shoot rooting, using DKW/gamborg and sucrose as the basal media for both. I still need to get some ppm, so I can minimize contaminates. Any recommendations on rooting? It seems like that is where things can get tricky. I also was spit balling the idea of adding a ppm or two i of nickel to the medium to help with nitrogen metabolism.

I expect to fail a lot, but it will be fun either way!

 

[englishrick]

That's exactly the paper I read ,,

Your on fire ,,,this is where you develop some special potions,,,

https://www.sciencedirect.com/science/article/abs/pii/S221478611530019X

It pains me to drop research in open fourm for companies to soak up , im sat in a shit legal climate and just can't do the work on my own to develop this shit properly

 

[englishrick]

I recently got prescribed medical cannabis here in the uk,,,the company is https://lyphe.com/ im considering asking for a job,,,back in 2010 i had started a pharmaceutical company but the world wasn't ready,,,medical cannabis was a pipedream at the time,,

The doctor told me to contact the team to look at being a consultant,,,anyone know who's running it??

 

[Dr.Mantis]

That's exactly the paper I read ,,

Your on fire ,,,this is where you develop some special potions,,,

https://www.sciencedirect.com/science/article/abs/pii/S221478611530019X

It pains me to drop research in open fourm for companies to soak up , im sat in a shit legal climate and just can't do the work on my own to develop this shit properly

Thanks Rick, that’s an awesome paper! I also completely understand about not wanting to divulge too much, for the exact reasons you stated. I think I have some polyploid GH cheese buds forming (will need to double check the stoma to confirm). If I do, I will start testing out things and keep the group up to date as I go along!

 

[LostTribe]

@englishrick et al Greetings!

This stuff is very interesting. Is there a book or site that has the basic info? Or recommended reading and resources? Is one of those hepa hoods required or can a simplified clean box suffice?

Once this is started can these be kept in stasis for storage? If so what environmental conditions are required? I read about artificial seeds wondering the same for that aspect?

The cleaning aspect seems like the place to begin reading. PM me with some reading recommendations! Following along!

 

[Rico Swazi]

Anyone incorporating activated charcoal in their media?
The paper I posted earlier (post 49) had this on the subject-

Various sterilization methods were tested to ensure shoot development from nodal explants by limiting the frequency of contaminating endophytes, which otherwise caused the death of explants. Seven commercially grown tetrahydrocannabinol (THC)-containing cannabis genotypes (strains) showed significant differences in response to shoot growth from meristems and nodal explants on Murashige and Skoog (MS) medium containing thidiazuron (1 μM) and naphthaleneacetic acid (0.5 μM) plus 1% activated charcoal. The effect of Driver and Kuniyuki Walnut (DKW) or MS basal salts in media on shoot length and leaf numbers from nodal explants was compared and showed genotype dependency with regard to the growth response.

The addition of activated charcoal appeared to improve growth on the MS medium; therefore, MS salts plus 1% activated charcoal was used in most of the experiments in this study. Activated charcoal, when added to tissue culture media, can absorb or bind some of the toxic waste compounds released from growing plants, in particular phenolic compounds, thereby improving in vitro plant tissue growth

Here's a how to MS , White, Gamborg, Heller et al

MS media (Murashige – Skoog) composition and preparation

Table Of Contents Introduction The composition of the growing medium is crucial in every experiment containing living organisms. The need for a well-balanced plant medium arose at the beginning of hydroponic cultivation and plant tissue cultures. The first recipes for plant tissue cultures are...

sharebiology.com

 

[LostTribe]

How long does it take to get rooted clones this way? I have been doing a great deal of reading this week on some of the basic aspects. Mainly components, media, still air box and kits.

The microcloner kit seems expensive when you can buy the media packets and vials seperately.

 

[Dr.Mantis]

How long does it take to get rooted clones this way? I have been doing a great deal of reading this week on some of the basic aspects. Mainly components, media, still air box and kits.

The microcloner kit seems expensive when you can buy the media packets and vials seperately.

You can buy everything you need to make media and vita/hormones much cheaper. The salts used in these media are quite cheap. Problem is, unless you wanna make 100L of media, measuring less than a mg of a micro is really tough. That being said, if you make the basal salts for 100l, then mix well, you can just take what you need from the bulk mix. You pay for convenience with the commercial stuff, but you can certainly do it all yourself if you are the tinkering type!

 

[englishrick]

Anyone incorporating activated charcoal in their media?
The paper I posted earlier (post 49) had this on the subject-





Here's a how to MS , White, Gamborg, Heller et al

MS media (Murashige – Skoog) composition and preparation

Table Of Contents Introduction The composition of the growing medium is crucial in every experiment containing living organisms. The need for a well-balanced plant medium arose at the beginning of hydroponic cultivation and plant tissue cultures. The first recipes for plant tissue cultures are...

sharebiology.com

If you look next to the far right jar I'm this picture you can see a tub of activated carbon,,,

I promise il explain everything I've been working on soon as I've got a chunk of money to put into this properly,,,

 

[nepalnt21]

shit, i hope the project is going well, this is a great thread.