Cloning Plants By Tissue Culture
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pharmacoping
Active Member
Soooooo cool nitro !!! my man !!!! I wish I was there to celebrate !! great photos !!
20% bleach for a0 minutes will kill the incised mj explant. 10% 10 minutes, rinsed twice,then 70% alcohol, couple minutes, rinsed again twice in sterile water. they dont die. they dont mold much. experiments in your environment, with plants cut from a different room than mine, may show different contaminations at different times I imagine. so probably personalized for each app. dirty plants? outside? inside? humid?dry? pets?pests? surely make huge differences in results all the way through.
Bonkleesha, do you have some ros pics in vitro, would be so cool here. most serious mj gardeners love flowers too !!
20% bleach for a0 minutes will kill the incised mj explant. 10% 10 minutes, rinsed twice,then 70% alcohol, couple minutes, rinsed again twice in sterile water. they dont die. they dont mold much. experiments in your environment, with plants cut from a different room than mine, may show different contaminations at different times I imagine. so probably personalized for each app. dirty plants? outside? inside? humid?dry? pets?pests? surely make huge differences in results all the way through.
Bonkleesha, do you have some ros pics in vitro, would be so cool here. most serious mj gardeners love flowers too !!
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heres some more info..
"When the plants start slowing in growth or the jar gets full of tissue, you'll need to "replate" or "subculture" (propagate) that tissue. Prepare your desired media and, in your sterile work area, remove the tissue from the jar it's growing in. If you simply need to replate (transfer the tissue to a new jar with fresh media without dividing), then just pull it out of the old jar and place it in the new media jar. If you need to divide and "subculture" the tissue, then you'll have to have a sterile work area to set it on so it can be divided or cut into pieces."
and again,,, there is so many different ways!!!EXPERIENCE rules
"When the plants start slowing in growth or the jar gets full of tissue, you'll need to "replate" or "subculture" (propagate) that tissue. Prepare your desired media and, in your sterile work area, remove the tissue from the jar it's growing in. If you simply need to replate (transfer the tissue to a new jar with fresh media without dividing), then just pull it out of the old jar and place it in the new media jar. If you need to divide and "subculture" the tissue, then you'll have to have a sterile work area to set it on so it can be divided or cut into pieces."
and again,,, there is so many different ways!!!EXPERIENCE rules
pharmacoping
Active Member
correct, but did it slow in growth? mine slow in growth 3 months later...just sayin, never a week. it will fill the jar with a ball of foliage(pics coming)
SO FOR ALL THE NEWBIES 
LIKE ME, THAT THINK YOU CAN DO IT YOUR SELF, YOU CAN,,, BUT THERE ARE DEFIANTLY SOME THINGS U CANT GET AROUND,,,, PLANT PRESERVE MIXTURE IS A MUST.NADCC/POOL SANITIZER WILL WORK FOR A SHOT TIME,,BUT PPM HAS A WIDER SCALE FOR KILLING BACTERIA,,,,BUT NADCC MIGHT BE BETTER FOR UR RINSING AND CLEANING,,IN PLACE OF BLEACH BECAUSE OF ITS ABILITY TO BE A PPM SUBSTITUTE,,,70% ALCOHOL IS STILL USED,,NADCC DOES NOT HAVE TO BE RINSED OFF,,BUT I USED LIKE A 10% OF THE DILUTE TO TRANSFER TO MY AQUARIUM LUB-BOR-A-TORY,,MIXES I USED:
--1/8 TEASPOON POOL/HOT TUBE SANITIZER
(DONT BREATH THIS AT ALL,,UR POT WILL TASTE LIKE CHEMICALS FOR DAYS,,SERIOUSLY ITS DANGER DANGER SKULLXBONES--CRUSH WELL,,POWDER "WELL" IT HELPS THE DILUTION
--FILL TO 750ML,,, SOMTHING WIDE YOULL NEED TO CRUSH THIS MORE,,TRY TO ALWAYS USE DISTILLED/COOKED WATER..
-THIS WILL BEE YOUR STUFF,,
DONT SMELL IT!!!!!!! ITS GOOD SHIT,, 
DONT WORRY!!--I USED UP TO 1/2TSP- /750ML AND RINSED,,THEY ARE CLEAN,,CLEAN OH AND ALIVE.
BUT USE 30ML/L OF UR SOLUTION TO 1000ML OF THE WATER,THIS WILL BE YOUR MEDIUM WATER..
!!!!!!!!!!BUT THIS WILL NOT BE AS GOOD AS PPM!!!!!!!
-FOR ALL OF YOU THAT JUST WANT TO PUT UR COOKER UNDER COLD WATER TO "COOL FASTER" NO NO NO NO...THIS "WILL" HAVE AN EFFECT ON THE CONTAMINATION ENTERING WITH THE COOLING PROCESS..SO JUST LIKE THE MAN WITH THE PICS TO PROVE SAID,,, JUST LET IT COOL ON THE STOVE TOP...
I DID THIS..... AND MOST-LIKELY HAD AN AFFECT ON THE LENGTH OF TIME BEFORE THE NADCC COULD'NT HANDLE IT.
IF UR NOT IN IT TO WASTE TIME,,,,,,U KNOW,,,, "TILL THE BOSS GATHERS WORK FOR THE SHEEP" TIME...OR WANT RESULTS FASTER,,,,,,, THEN,,,,,, MAKING A SQUARE WHEEL ROLL,,,WELL THEN,, THERES MORE "ON" THIS THREAD THEN SOME FLUNKY MATH,,,,,,CONSIDER IF THERE WAS A PERSON WHO ALREADY DONE THIS FOR ALONG TIME, AND HAD A KIT THAT WAS FOR SALE SOME WHERE ON THE NET. ..... PROBABLY WOULDNT GET MY HOPES UP FOR ANY INGREDIENTS TO THE MEDIUMS,,BUT,,WITH THE PICS TO PROVE/AND FOLLOWING DIRECTIONS, I WOULD TRUST THAT THIS DUDES KITS MAY WORK...I BET THAT GUYS CLOSE..SHIT HE MAY BE THE REASON MINE ARE STILL ALIVE AND GROWIN,,,, WITH PICS TO PROVE,,, HA HA!!!!!!
BUT I DIDNT BUY A KIT SO IF THIS IS WORKING,,,,LIKE I HEARD BEFORE,,,"JUST SAYIN!!"IF ANY "PROS" KEEP HEARIN THE SAME QUESTIONS OVER AND OVER THEN SEND THEM TO THIS LINK...IM SURE SOME WHERE IN HERE THEY WILL FIND IT!!!!! AND THERE WILL BE MORE TO COME FROM THE:
CULTURE GRASSHOPPER
LOVIN IT MAN,THANKS PHARMACOPING
pharmacoping
Active Member
wecome to the issue of Tissue my man !!
pharmacoping
Active Member
+ rep for the faucet and cool tile work on the counter....I'm gonna take a wild guess and suggest you did this yourself ?
pharmacoping
Active Member
heres some sultry culture snaps to drool over.... like I must have done on that moldy one. I 'm trying to capture the anomalies, rather than the normal ones. but the shooting/rooting pic is one I'll follow in this thread. shes strange as she got her shoot before her root. normally she would be culled from the herd, but for pix sake I'll keep her alive and fruiting.
I'll get some shots of rootballs and the transitions to veg, then flower soon. I am really really busy gearing up for the spring nursery rush, and cant take a camera in there, so my private tissue stashes are what you get , and in some sort of a time line for understanding.
the test tubes have agar in them mixed one month ago. none have molded, I'll keep them around for your entertainment. always used glass, but recently someone suggested these plastic alternatives. they suck, discarded after done, glass only man.
I'll get some shots of rootballs and the transitions to veg, then flower soon. I am really really busy gearing up for the spring nursery rush, and cant take a camera in there, so my private tissue stashes are what you get , and in some sort of a time line for understanding.
the test tubes have agar in them mixed one month ago. none have molded, I'll keep them around for your entertainment. always used glass, but recently someone suggested these plastic alternatives. they suck, discarded after done, glass only man.
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Bonkleesha
Active Member
heres a miniature rose... propagated on 1/25/12. i dont have pics from the other lab, but i have been doing photo documentation on this one 
Bonkleesha
Active Member
can we talk about the totiopotency of cannabis for a second? what is it comparible to? does indica or sativa have more? THESE are beneficial questions... anyone know how to get the answer?
pharmacoping
Active Member
both are equally totipotent in my experience
the pic is so cool....care to share your rooting formula for roses? and what you cap your tube with ? is it a glass tube?
the pic is so cool....care to share your rooting formula for roses? and what you cap your tube with ? is it a glass tube?
Git a light with a bad ass bulb,,had a little under the counter plant/aquarium light...now w should see some more growth....
pharmacoping,,is that electrical tape???? so what about test tubes with good rubber corks and then tape...grasshopper says,,i trust baby jars with the metal lid (dont have the plastic) taped,,into sandwich zip locks,,then tape the zippy part...but that dont mean nothin got in during trasfer,,,do it????NOPE..
SOURCES for plant preserve,,and supplies like forceps,,and glass?
oh!
pharmacoping,,is that electrical tape???? so what about test tubes with good rubber corks and then tape...grasshopper says,,i trust baby jars with the metal lid (dont have the plastic) taped,,into sandwich zip locks,,then tape the zippy part...but that dont mean nothin got in during trasfer,,,do it????NOPE..
SOURCES for plant preserve,,and supplies like forceps,,and glass?
oh!
nnnaaa,,,its came with the rental,,its a clean area to mix and cook...a glass pain is what i work off of..my work is outside and wondering if it should grow or not...this winter is WEIRD,,,everything has mildew,,and there is no snow..no ski park,,NO HOT ASS OUT OF TOWN SKI BUNNIES,,damn it...BETTER watch out for the mites in northern cali. this next summer....shit,,shouldnt trust a clone here if ur high depended on it..thats my experience..
Bonkleesha
Active Member
it was premade at some lab... the college orders em. im trying to get my hands on all the leftovers and even used ones with and without media they have had throughout the years.
i think its important to note that the cap is just thin plastic, but it goes over and around the tube, instead of in it like a cork. we made sure to sterilize the cap more than once, and we did it on the science lab under a high-suction HEPA hood. not the one with the enclosed gloves, but sterile enough. what i also want to get my hands on is the antioxidant that we soaked our explants in for 10 minutes directly prior to putting them into the media. do u want me to scan the lab instructions, dude?
oh and we just sealed the tube with some of that non adhesive plastic shit that stretches easy and sticks to itself.
i think its important to note that the cap is just thin plastic, but it goes over and around the tube, instead of in it like a cork. we made sure to sterilize the cap more than once, and we did it on the science lab under a high-suction HEPA hood. not the one with the enclosed gloves, but sterile enough. what i also want to get my hands on is the antioxidant that we soaked our explants in for 10 minutes directly prior to putting them into the media. do u want me to scan the lab instructions, dude?
oh and we just sealed the tube with some of that non adhesive plastic shit that stretches easy and sticks to itself.
pharmacoping
Active Member
. 10% bleach soak is standard in any culturing I've done in the nursery biz. thanks so much for the info...and stick aroundwtf?? of course I want the directions...what the heck was I thinking..sorrypost them or compress the or anything. you'll be sent love for any new info on the subject.
Bonkleesha
Active Member
how small would a cannabis explant have to be if you were going to microprop it to outgrow a disease?
pharmacoping
Active Member
I have no idea. I'm thinking that part is incidental for me, as I do this only to whip that plant count thing, without mothers even
Bonkleesha
Active Member
heres the info on definitions we got... im doing the rose lab now, and yes... the formula is one one of those pages (not these).
notice the antioxidant is plainly in view. i need to not go to class ripped :/
notice the antioxidant is plainly in view. i need to not go to class ripped :/
Bonkleesha
Active Member
heres the procedure and the formula for the media on the miniature rose. hope someone gets something out of it, if not, i just decided to scan all my handouts for easy reference in the future
