Cloning Plants By Tissue Culture
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wyteboi
Well-Known Member
well this is your thread now, the op seemed to dissapear an guile an myself are open ears. please maintain this thread too.
you know your tissue , an we need ya around. your a great help. thanks.
same with you guile , keep this thread up an going.
thanks guys!
soil
pharmacoping
Active Member
thanks for the kind words wyteboi ! I'm mixing up some custom mj rooting compound as we speak. it goes into vessels, sterilized,gelled, and shipped. the little mag stirrer is getting a workout today. I couldnt stand and mix for hours without my mind xploding !
pharmacoping
Active Member
"I keep seeing things that look more like what I would call "micro-propagation" than cell culture.. Maybe My impression of what "cell culture" might be is wrong... If you start with a seed or seedling it seems that you are only stretching the seed..I somehow figured we were looking to "clone" a mature plant using parts that would not otherwise be, like growing tips or other conventional clone/propagation material. More or less stems and/or roots..I was also under the impression that we were exploiting a survival/repair response from the plant to achieve this..An ultra simplistic perspective would be if you were to be able to say quarter a plants stem length wise to expose the Parenchyma, assuming its undifferentiated cells inside then influence it to spring up a column of miniature clones? Otherwise extract the undifferentiated cells and provoke them to make the callus that can be manipulated in that direction.If the Parenchyma of cannabis contains the undifferentiated cells cells you are after could the differentiated cells be dissolved or mechanically stripped away so that it were predominantly the undifferentiated cells being incubated? Would this provoke more successful callus formations?
You're way ahead of me there buddy ! I dont experiment much, not a mad scientist either.Short answer is....I have no idea,but heres what I do I take a cutting spec from a new tip, midway down. It grows callus, gets bigger. then I cut a piece off,transfer,and it grows roots. I transfer the roots, and they shoot up a bonzai tree quickly. I harden, then put them in the veg system. thats the extent of what I do today. I keep jars of "unusable roots" waiting for my order. anything done over the years was just figuring out what not to do, then directions came out to do this. I followed directions. on the cheap. with success. only issue is sterilizing plant material . the rest is second nature now.Only taking up a plant count number at the last moment when told to shoot, this allows me to have rooting genetics on standby, negating the seedling/cloning phase. these "seedlings" have a ball of roots that rival a month old plant....all before they even sprout !
Once a farm is established of chosen root ball genetics, and regular transfers take place, it really is a no brainer. lots of back ups in case of failures, common sense, years on the internet, or a week in a book. they're only super plants for a couple weeks, then look quite normal, except for the excessive lateral branching, ie lots of roots in the beginning.
many strains will grow from seed directly into flower(seed from 12/12) with minimal harvest weight loss. if i experiment any, it will be with moving a bonzai from a jar and putting directly in the flower room. other than that, I have no plans to exploit the technology any further than my own applications. lots of success can be achieved in vitro with a good rooting gel, on the cheap, if needed. varying amounts do really wild things in culture.
I culture plant material in different stages, so that I can micropropagate them to fulfill my needs. I hope that helps
You're way ahead of me there buddy ! I dont experiment much, not a mad scientist either.Short answer is....I have no idea,but heres what I do I take a cutting spec from a new tip, midway down. It grows callus, gets bigger. then I cut a piece off,transfer,and it grows roots. I transfer the roots, and they shoot up a bonzai tree quickly. I harden, then put them in the veg system. thats the extent of what I do today. I keep jars of "unusable roots" waiting for my order. anything done over the years was just figuring out what not to do, then directions came out to do this. I followed directions. on the cheap. with success. only issue is sterilizing plant material . the rest is second nature now.Only taking up a plant count number at the last moment when told to shoot, this allows me to have rooting genetics on standby, negating the seedling/cloning phase. these "seedlings" have a ball of roots that rival a month old plant....all before they even sprout !
Once a farm is established of chosen root ball genetics, and regular transfers take place, it really is a no brainer. lots of back ups in case of failures, common sense, years on the internet, or a week in a book. they're only super plants for a couple weeks, then look quite normal, except for the excessive lateral branching, ie lots of roots in the beginning.
many strains will grow from seed directly into flower(seed from 12/12) with minimal harvest weight loss. if i experiment any, it will be with moving a bonzai from a jar and putting directly in the flower room. other than that, I have no plans to exploit the technology any further than my own applications. lots of success can be achieved in vitro with a good rooting gel, on the cheap, if needed. varying amounts do really wild things in culture.
I culture plant material in different stages, so that I can micropropagate them to fulfill my needs. I hope that helps
Doer
Well-Known Member
Well, heat control is the reason you run 2 rooms and 2 lights, one ballast. And ballast can last longer if it's not cycled.
I've tried 8 weeks of cloning the traditional way and got Zero. And I have one of the WW in bloom that is a wonder grower compared to the other 4.
I'm contemplating a cloner. Invest $300. Surely this is more reliable and less expensive. Test tube environment is small and controlled.
pharmacoping
Active Member
I woud not suggest buying a cloner, if you're expecting different results. Rockwool, Replicator, Sharp Razor, angle cut mid section lateral shoot branch, scrape a little, dip, and place in the wet rockwool. follow directions on the rockwool. clones amore difficult to maintain if taken during a flowering cycle, but very possible. You can also harvest when ready, and leave some leafy small growth at the bottom attached, put it back in veg cycle, and you can try again.
tissue cloning will cost you more than 3 bills to begin. and it has to remain sterile. It cant replace traditional cloning for small grower, but is the only way to avoid using up clones as part of your legal plant limit.
tissue cloning will cost you more than 3 bills to begin. and it has to remain sterile. It cant replace traditional cloning for small grower, but is the only way to avoid using up clones as part of your legal plant limit.
Guile
Active Member
I probably should have read more before making speculations to start with. Alot of how I learn is to use my intuition (or incites based on other research/experiments) to speculate an outcome then pick up enough vocabulary and individual appreciation to move on and experiment.
I wouldn't go as far as to say that I'm a mad scientist, just learn best from first hand experiences.. Formulate a hypothesis, conduct experiments, record data (if you have the time, rinse and repeat). Then formulate a better educated theory/hypothesis and start again. At some point you will gain a true understanding of how many aspects of things conspire to obtain a particular outcome (now that's an education, understanding the Why as well as the how). Honestly its how I always learned, my mother tells a story of me disassembling spring loaded cupboard hinges (child safety hinges) to see how they worked back when I was 3.
Knowledge is the product of Knowing, how can you know if you never did? Otherwise you are just accepting other peoples individual appreciations (which were likely guided by a paper written by someone else, subject to their own individual appreciations). as absolute fact. (where I accept my own bias, I sure as hell am not going to exclude anyone else's).
I do hope you realize how much I appreciate the information and incites you share. I've just been listening and reading a little bit more lately because at this point I already have alot to process before moving much further forward (otherwise I might lack fundamental understandings required to build from)..
I wouldn't go as far as to say that I'm a mad scientist, just learn best from first hand experiences.. Formulate a hypothesis, conduct experiments, record data (if you have the time, rinse and repeat). Then formulate a better educated theory/hypothesis and start again. At some point you will gain a true understanding of how many aspects of things conspire to obtain a particular outcome (now that's an education, understanding the Why as well as the how). Honestly its how I always learned, my mother tells a story of me disassembling spring loaded cupboard hinges (child safety hinges) to see how they worked back when I was 3.
Knowledge is the product of Knowing, how can you know if you never did? Otherwise you are just accepting other peoples individual appreciations (which were likely guided by a paper written by someone else, subject to their own individual appreciations). as absolute fact. (where I accept my own bias, I sure as hell am not going to exclude anyone else's).
I do hope you realize how much I appreciate the information and incites you share. I've just been listening and reading a little bit more lately because at this point I already have alot to process before moving much further forward (otherwise I might lack fundamental understandings required to build from)..
pharmacoping
Active Member
Hey Guile,
Thank you for the kindness share. I have to assume that you spelled "incites" rather than "insights" purposely. now thats wit !
Thank you for the kindness share. I have to assume that you spelled "incites" rather than "insights" purposely. now thats wit !
Guile
Active Member
No problem...
If you scrutinize my writing enough you would likely find many spelling errors. I'm a phonetic speller for the most part, though I use spell check to help identify where communication issues might arise (the web browser plugin I use has its limitations). Despite having been enrolled in multiple typing/keyboarding classes I'm not much of a typist either.
I use to have a hard time dealing with my lack of spelling prowess, particularly in elementary school (the level of education that they seem to focus on that sort of thing). I was actually labeled as "learning disabled" in 1st grade due to my lack of intuition when it comes to spelling. Ironically or perhaps more serendipitously I also ended up in the "gifted & talented" programs as well. My pear group growing up was a product of the polar ends of the educational standard. It was a bit of a culture shock when I went away to collage at 16 (I had received a full academic scholarship when I was 15 however the law required I become 16 before leaving high school). As I high school dorp-out (do to never legitimately passing an English course) I made my collage eligible for some kind of government program because I was a "minority"..
To be honest I'm socially awkward too... People rarely understand me well, the way people seem to react to my words leaves me to believe that many times there is a complete mistranslation of my intent or appreciations. Much like everything I do, I use words intuitively, biased on the "feeling" I get (gut instinct so to speak).
I never realty thought/cared much for my mind (and the way it worked) until I suffered a traumatic brain injury a few years ago.. Early on during my lengthy stay in rehab I underwent a bit of mental testing. After the first battery of tests the doctor told me that I could easily be considered "normal" given my background and education (I've gone to a couple different schools).. And that I had retained at least 95% of my mental capacity. Considering how "stupid" I had been up to that point I decided to take whats left of my mind more seriously and not waste it.. (if you don't use it you loose it).
If you scrutinize my writing enough you would likely find many spelling errors. I'm a phonetic speller for the most part, though I use spell check to help identify where communication issues might arise (the web browser plugin I use has its limitations). Despite having been enrolled in multiple typing/keyboarding classes I'm not much of a typist either.
I use to have a hard time dealing with my lack of spelling prowess, particularly in elementary school (the level of education that they seem to focus on that sort of thing). I was actually labeled as "learning disabled" in 1st grade due to my lack of intuition when it comes to spelling. Ironically or perhaps more serendipitously I also ended up in the "gifted & talented" programs as well. My pear group growing up was a product of the polar ends of the educational standard. It was a bit of a culture shock when I went away to collage at 16 (I had received a full academic scholarship when I was 15 however the law required I become 16 before leaving high school). As I high school dorp-out (do to never legitimately passing an English course) I made my collage eligible for some kind of government program because I was a "minority"..
To be honest I'm socially awkward too... People rarely understand me well, the way people seem to react to my words leaves me to believe that many times there is a complete mistranslation of my intent or appreciations. Much like everything I do, I use words intuitively, biased on the "feeling" I get (gut instinct so to speak).
I never realty thought/cared much for my mind (and the way it worked) until I suffered a traumatic brain injury a few years ago.. Early on during my lengthy stay in rehab I underwent a bit of mental testing. After the first battery of tests the doctor told me that I could easily be considered "normal" given my background and education (I've gone to a couple different schools).. And that I had retained at least 95% of my mental capacity. Considering how "stupid" I had been up to that point I decided to take whats left of my mind more seriously and not waste it.. (if you don't use it you loose it).
pharmacoping
Active Member
a kinetic personality for sure, is the most intense, and misunderstood, in my experience. I hope we have many years to share.
peace
peace
and the there is this. this i found last night and sums up tc for mj for dummies like me.
http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf
http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf
pharmacoping
Active Member
nitro my man ! gee i'm still wondering why an expensive lab would waste time culturing marijuana !!!!!
MONEY MONEY MONEY, this is "THE CASH CROP".yeah i wonder if the funding was payed in full by results of EXCELLENT CANNABIS??? k so heres another chemistry question, if i would have listened in school i probably would know this, or at the very lest at 36 yrs. old,,,forgot it.......ok so here it is,,,if i have a concentration of iba 1.0% & 0.5% naa,,,,to get to 0.1 iba & .05 naa mg./l??????
today is the day,, up until 1:00 a.m. cleaning and building.oh and thinkin.got to stop that.
my friend that has been doin this turn around on the mj tree $$$$ thing has mite and white mold. his mothers (king kush, grand daddy purps,and of course my poor blue dream). getting hard to clone and easily pics up root rot etc.... perfect, that is one of the reasons to do this,, ill fix em'!!!!!!!!
today is the day,, up until 1:00 a.m. cleaning and building.oh and thinkin.got to stop that.
my friend that has been doin this turn around on the mj tree $$$$ thing has mite and white mold. his mothers (king kush, grand daddy purps,and of course my poor blue dream). getting hard to clone and easily pics up root rot etc.... perfect, that is one of the reasons to do this,, ill fix em'!!!!!!!!
Guile
Active Member
MONEY MONEY MONEY, this is "THE CASH CROP".yeah i wonder if the funding was payed in full by results of EXCELLENT CANNABIS??? k so heres another chemistry question, if i would have listened in school i probably would know this, or at the very lest at 36 yrs. old,,,forgot it.......ok so here it is,,,if i have a concentration of iba 1.0% & 0.5% naa,,,,to get to 0.1 iba & .05 naa mg./l??????
today is the day,, up until 1:00 a.m. cleaning and building.oh and thinkin.got to stop that.
my friend that has been doin this turn around on the mj tree $$$$ thing has mite and white mold. his mothers (king kush, grand daddy purps,and of course my poor blue dream). getting hard to clone and easily pics up root rot etc.... perfect, that is one of the reasons to do this,, ill fix em'!!!!!!!!
If you simply want to dilute the concentration 10:1 then you just want to add 9 volumes distilled water to 1 part concentrate..
If you are trying to obtain 0.1 ppm from a solution of 1% Iba the easiest way would be to try to find something that mentions PPM in its dilution ratios (then just scale accordingly).
Otherwise whats it's specific mass/density? (the figure listed on its chemical, MSDS, or CAS description) also is it 1% by mass or 1% by volume? (to establish a ratio biased no mass/density). If your base solution has a different specific gravity than water you may also have to take that into consideration (depending on how much information you can come up with and how you want to skin that cat).
its cool what you can figure out with a scale, graduated measure, hydrometer and a calculator (if your resourceful you might get away with half that stuff.) Math is beautiful dude and its a language that's easy to respect (not alot of special rules or contradictions).
man i love this. yeah i got the conversions and 1 more page of notes.
but i dont have an equation to solve, ur right,,,so help skin this cat(hate cats) dip n grow is a alcohol base liquid and it is 1.0%iba & .5%naa by wieght,,98.5% other ingredients.smells 100% alcohol. so, with that i want to get it to .1mg/L iba..it comes with a concentration cup,,1 tsp. is the X line 5x = 5 tsp. i guess that was obvious. the naa will be what its is.Im shooting for the #2test result of this study:http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf
Guile
Active Member
98.5% other ingredients does pose a bit of a problem... Can you get a breakdown of the other ingredients and ratios they are used in?man i love this. yeah i got the conversions and 1 more page of notes.but i dont have an equation to solve, ur right,,,so help skin this cat(hate cats) dip n grow is a alcohol base liquid and it is 1.0%iba & .5%naa by wieght,,98.5% other ingredients.smells 100% alcohol. so, with that i want to get it to .1mg/L iba..it comes with a concentration cup,,1 tsp. is the X line 5x = 5 tsp. i guess that was obvious. the naa will be what its is.Im shooting for the #2test result of this study:
http://www.pakbs.org/pjbot/PDFs/41(2)/PJB41(2)603.pdf
I would guess that you might want to find out the specific gravity of the solution (with a hydrometer) then the specific mass, derive the mass of the 2 active ingredients (1.5% @ the appropriate ratios and quantity biased on weight). From their chemical descriptions you should be able to determine their mass per volume (specific density) now I think you can figure out the ppm biased on mass, specific density, and molecular weight. I'm sure you can find a calculator for it online unfortunately I don't know the formula.
I'm also high and despite rewriting this like 3 times it still doesn't sound right to me, but you are only figuring out how many parts are in a volume biased on weight (specific mass) and applying it to the ratios established on the label (also biased on weight) to determine the number of parts in the mass of hormones used (for the given volume of concentrate).
The molecular weight/specific density (mass per volume) of the hormones are likely the key considerations...
Here is a simple one :One ppm is equivalent to 1 milligram of something per liter of water (mg/l) or 1 milligram of something per kilogram soil (mg/kg).
So essentially, 1ppm = 1mg/l Source(s):
http://groups.molbiosci.northwestern.edu/holmgren/Glossary/Definitions/Def-P/parts_per_million.html
It doesn't take into consideration the unknown specific gravity of your base solvent but I doubt you would be off by much..
1 liter of water would weigh 1000grams 1% of which would be 10 grams, if 1mg/L = 1ppm, then 10000 milligrams should equal 10,000ppm (that seems like alot to me) what ratios do you mix this stuff for say foliage feed or long soak application?
yep.,,,math and numbers ,,theres somthing great about it all..i guess it would be the oldest universal language know to animals with a propose.(man kind)k,,,so this is what i needed to get to a ppm??http://www.dipngrow.com/wp-content/uploads/2010/08/Parts-Per-Million-PPM.pdfwell here it is now i got it. but im still not done solveing the last equation....so weight by mass of iba=A weight solution (dip and grow) =X i think its divide A into X = W there it is.i think. then convert W to mg/L. To think i would pay 1000.00s to sit and be graded on this weeks learning disability.this is way funner.
OK TISSUE CULTURE DUDES u can have ur site back, now its time for results WITH PICTURES i know.
Guile
Active Member
It looks like your dip n grow would be proximately 10,000 ppm IBA and apx 5,000ppm NAA. Assuming Dip n Grow isn't flammable at room temperature, its base solvent is predominantly water and those figures should be within 10% accurate.
so if you wanted a concentration of 0.1ppm you would use a ratio of 1:100,000 or 1ml to 100L..
I would probably go about it by putting 1ml Dip n Grow into a liter container and top off with water to get 10ppm then take 10ml of that solution and dilute into another liter of water to get 0.1ppm.
I'm a bit more sober now and it still feels wrong but I'm not seeing where I'm messing up yet...
Don't worry too much about my earlier ramblings, I was high and over complicating things. this equation for finding ppm biased on mass and the density of water should be within 10 percent (or so) assuming that Dip'n Grow is under 100 proof..
pharmacoping
Active Member
whoaaa, you a smarty pants guile ! thanks man, good to see a serious mind here.
I buy it already diluted, because I never learned how to drive a calculator
I buy it already diluted, because I never learned how to drive a calculator
yep should have went with a less concentrate. newbie's!!! think that we can do cheaper/better.no you are right with that one. Thats how i was thinkin it was broke down.. but listen to this one: NxE=JUSTBUYTHEKIT with a remainder of PRESURECOOKER!!!ha aha ha haha. Newbie x Experience= always a realization.but back to work (with pictures i know)i think i have to re-size them cause that firewall thing..connection..etc. would let this old ass Dell through.lights closet,hepa running now,bleach n clean. to ace for plastic(mini lab)powered gloves no good. to my buddy's ,oshit things to do, cant rush this.LATER!!!oh goin with @1000ml-1 to 1.5 ppm iba, which will make ppm naa @ .5 lower,,perfect!!!