New Stock
- Thread starter PANIKZ
- Start date
For those of you messing with tissue culture and older seed:
Tween 20
Zerotol
Agra-Rouse
All things you should have on your shelf.
dank.Frank
can u explain how they are each used. i saw first 2 are like anti pathogen/spores etc.
Old seeds is my issue and an edge that's economical that makes a difference is worth trying.
i saw today distilled water in 1/2 red cup, hydrogen peroxide and 1/2 tablespoons organic coconut water . not kidding.
Detailed method on germinating old seed
Quote:
Originally Posted by Chimera
The protocol I use is pretty simple.
5% bleach in autoclaved, distilled water with a couple of drops of Tween-20 per liter of solution. I sterilize the water in Duran jars (loose lids) in a pressure cooker @ 20 PSI for 20 minutes, and then allow the jars to cool under my flow hood and tighten the lids when the water has cooled. This is probably overkill for most purposes, but it's a variation of the protocol I use for sterilizing explants for tissue culture, and it seems to work with minimal contamination. You can get away with a drop of dish detergent instead of the Tween, I just have the tween-20 in the lab so I use it. If you are going to be doing a lot, you can order a few ml of Tween-20 on Amazon if you like.
First I rinse the seeds in a tight mesh strainer to remove any leaf debris that might be harboring bacteria or spores. These seeds were a little dirtier than most having been field grown, so you definitely want to wash away any superfluous materials. This can be up to a 5 minute rinse in lukewarm water. Cannabis achenes have a lot of little crevices where spores and bacteria can hide, so a thorough rinse is a wise step.
Then I take the seeds and drop them in maybe 100-200 mls of the bleach/Tween-20 solution in an Erlenmeyer flask, but any clean glass vessel will do -the seeds should be fully immersed, maybe 2x the volume of water as seed. You want to keep stirring and swishing the seeds around gently to ensure the current is washing away any contaminants. After a minute of swishing I remove the bleach and strain the seeds from the liquid and start again. No more than 3 minutes total, and if I see bleaching on the small test batch (always do a test batch to seed how the seeds react), sometimes I'll only do 2 minutes.
The next part is key. Immediately rinse 3-5 times (for 5 minutes per rinse) in the same sterilized distilled water (baby water / distilled water from Walmart or the local pharmacy is fine), again using a tight mesh strainer, or simply rinse the seeds in a different vessel with the same volume of water, at least 3 times to ensure the bleach is removed. I use a labratory orbita shaker, but you can do it by hand it's just a little tedious. Once the rinses are completed, soak the seeds in the distilled water as before, for 2-24 hours depending on the age of the seed and the medium you will be growing in. Sometimes I'll use agar plates, but most often I'll just sprout them in paper towels and transfer to a light dirt (like Light Warrior for you Americans) or surface sow on a .peat based planting substrate. You can use whatever medium you prefer, it's not that big of a deal as long as the seeds remain moist
Quote:
Originally Posted by Chimera
The protocol I use is pretty simple.
5% bleach in autoclaved, distilled water with a couple of drops of Tween-20 per liter of solution. I sterilize the water in Duran jars (loose lids) in a pressure cooker @ 20 PSI for 20 minutes, and then allow the jars to cool under my flow hood and tighten the lids when the water has cooled. This is probably overkill for most purposes, but it's a variation of the protocol I use for sterilizing explants for tissue culture, and it seems to work with minimal contamination. You can get away with a drop of dish detergent instead of the Tween, I just have the tween-20 in the lab so I use it. If you are going to be doing a lot, you can order a few ml of Tween-20 on Amazon if you like.
First I rinse the seeds in a tight mesh strainer to remove any leaf debris that might be harboring bacteria or spores. These seeds were a little dirtier than most having been field grown, so you definitely want to wash away any superfluous materials. This can be up to a 5 minute rinse in lukewarm water. Cannabis achenes have a lot of little crevices where spores and bacteria can hide, so a thorough rinse is a wise step.
Then I take the seeds and drop them in maybe 100-200 mls of the bleach/Tween-20 solution in an Erlenmeyer flask, but any clean glass vessel will do -the seeds should be fully immersed, maybe 2x the volume of water as seed. You want to keep stirring and swishing the seeds around gently to ensure the current is washing away any contaminants. After a minute of swishing I remove the bleach and strain the seeds from the liquid and start again. No more than 3 minutes total, and if I see bleaching on the small test batch (always do a test batch to seed how the seeds react), sometimes I'll only do 2 minutes.
The next part is key. Immediately rinse 3-5 times (for 5 minutes per rinse) in the same sterilized distilled water (baby water / distilled water from Walmart or the local pharmacy is fine), again using a tight mesh strainer, or simply rinse the seeds in a different vessel with the same volume of water, at least 3 times to ensure the bleach is removed. I use a labratory orbita shaker, but you can do it by hand it's just a little tedious. Once the rinses are completed, soak the seeds in the distilled water as before, for 2-24 hours depending on the age of the seed and the medium you will be growing in. Sometimes I'll use agar plates, but most often I'll just sprout them in paper towels and transfer to a light dirt (like Light Warrior for you Americans) or surface sow on a .peat based planting substrate. You can use whatever medium you prefer, it's not that big of a deal as long as the seeds remain moist
Zerotol sanitizes everything. Tween 20 helps penetration of materials and even coverage. Agra-Rouse is a brilliant combination of PGR's which are typically used individually for stimulating growth and cell elongation.
That aside, simple sugar water is also another thing used during invitro germination to stimulate growth. Makes sense. Glucose is part of the ATP cycle.
dank.Frank
That aside, simple sugar water is also another thing used during invitro germination to stimulate growth. Makes sense. Glucose is part of the ATP cycle.
dank.Frank
Hi mate I have Blue Satellite, Bluebonic, an Donk viability is awful I’d imagine as these are old but shoot me a pm we can arrange some at a bargain price , did you message via another forum about these ???
Greatdalas
Well-known member
I'm interested In Dutchgrown seeds (thai dragon, Xanadu and others)
Yes have loads of these mate ..
Greatdalas
Well-known member
Can you seid me how much is the packets of Dutchgrown and it is good the germination or don't known, thanks
I would think the viability isn’t great ..
Pm me mate we can sort out a deal on them
Greatdalas
Well-known member
Mullumbimby madness x nevilles haze 21 they germinated all
2 famales
1 hermie
And the other males
Are Medium sized plants
2 famales
1 hermie
And the other males
Are Medium sized plants
wow mate they look lovely , what stock are they from the Spice of life stock we have ???
Earlier you mentioned O. Haze x love potion x skunk/haze,
O. Haze / skunk x love potion x haze/skunk.
How are those coming along?
of course, anyway i wouldn t mention it as "same stock"
Greatdalas
Well-known member
Are a good strain, germinate all and have very much phenotypes
Appreciate the feedback, thanks
Did you finish flowering? How long did they go indoors?
Greatdalas
Well-known member
Appreciate the feedback, thanks
Did you finish flowering? How long did they go indoors?
No, have 30 days flowering now
H x s/lp/h x s
H/lp/h x s
Hawaiian indica/h x s
