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Cheap Tissue Culture

  • #61
It's cool man. I did like the comment about the colors, I didn't know mold came in so many forms.

@BigBella: Interesting. Are there any vitamin pills that do have something similar to a PGR in them that you know of? I'm just curious, for now I think I am done buying materials.
 
  • #62
There are related zeatin and purine supplements, which are readily available. Among the earliest of the PGRs, kinetin, a specific purine, was first extracted from spoiled herring semen. I don't even want to hazard a guess what Miller and Skoog did in their off hours; or, more significantly, how they determined that the semen had turned . . .
 
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  • #63
LOL (I mean it)
 
  • #64
word around town was that Folke would walk the docks looking for the freshest specimen...
 
  • #65
:lol: Anything for science I guess. I did do a search for kinetin and zeatin supplements, and found this http://www.amazon.com/Kinerase-Ther...ie=UTF8&qid=1363450934&sr=8-1&keywords=zeatin. Hard to administer, though, and costs twice as much as this whole project combined.

EDIT: I read a few papers, and it appears that seaweed extract also contains many cytokinins. One abstract of a study concerning commercial brands said that "Cytokinin activity of the seaweed extract was confirmed by the increase in growth it produced with tissue-cultured cells...The results obtained show that the seaweed extract has a cytokinin activity capable of producing physiological changes, even when applied at the low concentrations used in practice" (Brain). Maybe something like this could work? http://www.amazon.com/Maxicrop-Liqu...id=1363455273&sr=8-2&keywords=seaweed+extract Or maybe this? http://www.homedepot.com/p/t/203615...nosearch3-5-_-NA-_-203615935-_-N#.UUSuQ1vwIeA
 
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  • #66
I limited the earlier response to the question of supplements; but the use of all manner of foodstuffs have been attempted and shown to have some hormonal activity or influence growth, including orange juice, for example. It can promote callus growth in Citrus species, though it is thought to be the influences of organic acids more than anything else; and banana homogenates have been used for decades in orchid culture; though, whether it is the presence of PGRs in some of the substances; an aid to the Kreb Cycle, associated with organic acids; or, simply the stabilization of in vitro pH, thought to be associated with banana; the jury is still out . . .
 
  • #67
That's really cool. So maybe I'll add a smoothie to the mix? :lol:
 
  • #68
I just finished my first test with the pressure cooker. I pressurized three different baby food jars with different amounts of cornstarch for 15 minutes to see how it gelled. It went remarkably well (until I dropped one of the jars :(). The jar with 6 teaspoons of cornstarch per 1/2 a cup of water gelled perfectly, so I think that is what I will use with the real deal tomorrow. I also bought some neon food coloring, so now it's a real mad scientist experiment!


6 teaspoons per 1/2 cup of water:

<a href="http://www.flickr.com/photos/84442298@N03/8562369027/" title="DSC02214 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8520/8562369027_64ef002181_z.jpg" width="640" height="480" alt="DSC02214"></a>


5 teaspoons per 1/2 cup of water:

<a href="http://www.flickr.com/photos/84442298@N03/8563476868/" title="DSC02213 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8392/8563476868_f46187890e_z.jpg" width="640" height="480" alt="DSC02213"></a>


4 teaspoons per 1/2 cup of water:

<a href="http://www.flickr.com/photos/84442298@N03/8562369049/" title="DSC02212 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8530/8562369049_1a840565ce_z.jpg" width="640" height="480" alt="DSC02212"></a>


Having a pressure cooker on high heat for too long is one of the scarier experiences in life...
<iframe src="http://player.vimeo.com/video/61972117" width="500" height="281" frameborder="0" webkitAllowFullScreen mozallowfullscreen allowFullScreen></iframe> <p><a href="http://vimeo.com/61972117">Pressure Cooker</a> from <a href="http://vimeo.com/user17108509">Drew E</a> on <a href="http://vimeo.com">Vimeo</a>.</p>
 
  • #69
I'd advise avoiding any food coloring in the media, until you get a handle on contamination -- which is a far greater concern. Colorings are generally added to code for specific media and aren't necessary for the time being.

Oh, and stop dropping things . . .
 
  • #70
I thought that food coloring could help pick out contamination? Or does it spread as the same color?

I'll try. Baby food jar lids have very few threads. 10 left to experiment with.
 
  • #71
Sorry for another double post, but it is unrelated to the above. In reading about cytokinins, it appears that they are produced by the root tips of a plant in excess; roots supply all of these hormones for the entire plant. Do people use root tips in culture? Are they very difficult to sterilize?
 
  • #72
Some suggestions:
• Fertilizer: choose fertilizer with completed macro and micronutrient (N,P,K,Ca,Mg,Mo,Zn,Mn,Cu,Fe) Try to avoid fertilizer with urea because heat may convert urea to harm chemical.
• Buy agar pouch at Asian market for $1.50.
Agar_front.jpg


• Use pickle/jam/pepper … jars from dollar store, 1 jar 1 dollar. They look good, cheap and much better than baby food jar.

jars.jpg


• You need a digital 100g x 0.01g balance from Ebay for less than $10.
• Now, it's time for practice, try seeds first: sterilize and sow seeds. :-D
 
  • #73
Thanks for all the advice. What else do you add to your media besides fertilizer and agar? At the moment I have 10 jars that were given to me for free, but I will certainly check out the dollar store in the future. Thanks for your advice!
 
  • #74
Basic: light + temperature + agar + sugar + fertilizer
Google for “The Components of Plant Tissue Culture Media I Macro & Micro” for more information about macro and micro nutrient.

Enhance: vitamin, inositol, potato/banana/tomato extracts, amino acids, organic acids …
Google for “The Components of Plant Tissue Culture Media II organic addition”; review it, compare with MS media, and add supplements if you can to improve your medium.
 
  • #75
@Flasker: You can see the medium I use in the previous page of this thread. Thanks for the suggestions, I'll read those papers for my next attempt.

WOOHOOO!!! I completed the first five jars of media today. I took out half a cup of the liquid and added 6 teaspoons of cornstarch. I used a syringe to transport about 30mL to every jar. I boiled them in the pressure cooker for about 30 minutes, and it all went without a hitch. One jar opened when I was trying to pull it out, which is unfortunate, but I closed it as quickly as possible. All of the jars looks great, there was some stray liquid in one or two jars, but since cooling they have solidified completely. They are currently Saran-wrapped in my basement awaiting cuttings (probably planted by Tuesday). I'm so excited. On my blog I will have a video quickly showing the process up by Tuesday.


Pics:

The whole batch.

<a href="http://www.flickr.com/photos/84442298@N03/8566175853/" title="DSC02225 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8088/8566175853_cf63f47db5_z.jpg" width="640" height="480" alt="DSC02225"></a>


With flash.

<a href="http://www.flickr.com/photos/84442298@N03/8567272004/" title="DSC02227 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8521/8567272004_31e259b776_z.jpg" width="640" height="480" alt="DSC02227"></a>


The real color without flash. Not that gross!

<a href="http://www.flickr.com/photos/84442298@N03/8567271828/" title="DSC02228 by Sundrew, on Flickr"><img src="http://farm9.staticflickr.com/8103/8567271828_47ed67283b_z.jpg" width="640" height="480" alt="DSC02228"></a>
 
  • #76
You gonna start selling these? Or should we just send you the cuttings/seeds? :awesome:
 
  • #77
The jars remind me of an odd dessert I had in a Asian restaurant -- sort of a pudding made with agar with almond flavoring.

How does the media taste?
 
  • #78
LOL if I had the experience to tell you I'd be knocked out on the floor from the ferts...

Someone else told me it looked just like where Miller and Skoog first found cytokinins.
 
  • #80
LOL

Someone else told me it looked just like where Miller and Skoog first found cytokinins.

Jello® Spoiled Herring Semen Parfait?
 
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