Using genetics to determine a plants sex

[thegenemaster]

Hi all, this is my first post. I'm actually a scientist. I currently do cancer research but I had been a plant molecular geneticist at one point in my past. I started growing medical cannabis a couple years back and wondered if it would be possible to determine the sex of a plant simply by looking at the genes. In this way you could identify males and females as very small seedlings. Bottom line is I have done it. I use a marker to screen my plants and separate out the males.

I developed this technique just for my own purposes but now have been told that a great many growers would really benefit from my service. It has been tried on over fifty varieties and identifies the males in everyone tried to this point. I guarantee it works. I am set up to analyze a lot of plants and I can do it for a very reasonable price. Please give me some feed back on this. Thanks!

[stinkyattic]

If this is a sales post, it's in violation if site rules. Please read the forum guidelines concernig unauthorized advertising, and contact the site admin if you would like to advertise legitimately, thanks! :)

Sex in cannabis is 'set' during the very early seedling stage. You can easily force 99.9%+ female by simply pollinating a 'normal' female with pollen from a female that has, either naturally or through photoperiod manipulation or even chemical treatment, produced male sex parts. These 'nanners', as growers call them, are interesting in that, while they produce pollen that by all appearances is in fact viable pollen, it is physically impossible for it to contain a Y chromosome. Therefore, all offspring will be female.

Google 'feminizing seeds' and you will eventually find an interesting article by the breeder Soma on this subject.

[Weedhound]

Go Stinskter!! :)

[GreenLeaf420]

I guess it is easier to find the sex of a plant then to find a cure for cancer....

If you don't want to share then you must not have been around this site to long. If you understand the whole theory of the site it is to HELP others not to sell your services.

So if you wont tip us off hurry the fuck up and find the cure for cancer. We have been sexing plants longer then cancer has been killing hundreds of thousands people. We do our job so you do yours ,shit you are probably typing @ work and the cure for cancer could have just passed you by.....:mad:

I don't believe you are a scientist. Do you know what plant grows in HAWAII and cures skin cancer? Answer that and I will believe you....;)

[Forwhat420]

Well Im just wondering...How this info is useful? If one wants all females just buy them that way, otherwise some of us like the process of guessing and then waiting to see if we are right. Anywho I don't think anyone wants to buy that idea on the site. But if you want to give it to us then shoot!!

[thegenemaster]

I'm happy to tell eveyone how to do it. Use one of the half dozen methods available to extract DNA for a piece of plant tissue. I recommend using younger tissue as it has more cells per unit area. Determine the relative concentration of DNA and adjust it to approx. 50ng/ul(you will need a refrigerated centrifuge for this). Set up PCR(Polymerase Chain Reaction) each reaction should contain aproximately 50ng of sample DNA, 1x Taq polymerase reaction buffer, I recommend using 1.5mM MgCl2 but this may vary depending on the specific Taq polymerase you use. Aslo include in your reaction mixture approx. 0.02Units of Taq(thermostable DNA polymerase). Finally you will need to include a forward and reverse primer for one of the five known molecular male markers for cannabis. The concentration of these will need to be determined empirically as the specificity of the reaction is determined by the specific primer set you elect to use.

Now your ready to run your reactions. Be sure to keep the samples on ice prior to running the reactions. Place your samples in a previously calibrated thermocycler. These are available from a variety of vendors and a reasonably priced one is about 15K. Okay if you have made it this far contact me and Ill tell you how to set up your specific program interms of annealing Tm and hold times and recommended number of cycles to avoid non-specific amplification. When you contact me please be sure to let me know what the Tm is of each of your specific primers.

Okay now run your reaction out on a 1% 1xTBE agarose gel.

[thegenemaster]

Run the gel for about an our at 100v using an appropriate power supply. Now stain the gel with ethidium bromide, place the gel on a uv light box and take a photograph . Your done.

Look obviously I can tell you all how to do it but it just so happens I have all the equipement supplies and knowledge to to this and have done it several times.

I didn't develop this to make $$ I developed it to save my self tremendous time energy and money as a grower. Friends said there would be alot of people that would benefit form this service and would be happy to pay a reasonable fee for the service. Obviously I'm not going to do it for free.

I am a very capable of sexing plant as are many of you. But a can any of you do it before the plant even has true leaves???

[midlifecrisis]

Damn!!!!! My $25,00 dollar thermocycler is in the shop..........Oh well, guess I'll just have to order some feminized seeds again.....:jointsmile:

[thegenemaster]

I'm not trying to get rich.. and yes I have a fucking day job. Man I have just come this form this morning and I have apparently hit the asshole motherload.

FYI I am a very very good scientist. My research has directly led to the development of a drug that is scheduled to go into clinical trials within the year. I have been in the field for better than 15 years and less then 0.1% of any cancer researchers can make that claim.

So again, I'm only here to lend my expertise to fellow growers if they so desire it. Sorry I can't do it for free but I'm not going to go broke helping others out. Thanks for listening and in the future a little more decorum to fellow medical cannabis users and grows new to the forum is in order.

[thegenemaster]

available for you in every variety your interested in growing? And I have made and used femenized seeds in the past and my experience is that they have less than optimal genetic background.

[GreenLeaf420]

I'm happy to tell eveyone how to do it. Use one of the half dozen methods available to extract DNA for a piece of plant tissue. I recommend using younger tissue as it has more cells per unit area. Determine the relative concentration of DNA and adjust it to approx. 50ng/ul(you will need a refrigerated centrifuge for this). Set up PCR(Polymerase Chain Reaction) each reaction should contain aproximately 50ng of sample DNA, 1x Taq polymerase reaction buffer, I recommend using 1.5mM MgCl2 but this may vary depending on the specific Taq polymerase you use. Aslo include in your reaction mixture approx. 0.02Units of Taq(thermostable DNA polymerase). Finally you will need to include a forward and reverse primer for one of the five known molecular male markers for cannabis. The concentration of these will need to be determined empirically as the specificity of the reaction is determined by the specific primer set you elect to use.

Now your ready to run your reactions. Be sure to keep the samples on ice prior to running the reactions. Place your samples in a previously calibrated thermocycler. These are available from a variety of vendors and a reasonably priced one is about 15K. Okay if you have made it this far contact me and Ill tell you how to set up your specific program interms of annealing Tm and hold times and recommended number of cycles to avoid non-specific amplification. When you contact me please be sure to let me know what the Tm is of each of your specific primers.

Okay now run your reaction out on a 1% 1xTBE agarose gel.

What ever Mumbo Jumbo you are speaking up there may or may not mean anything.

Why would anyone consider to go through all those mesures to find out the genetics of one seed?

If you had any type of buisness sense you would understand COST EFFECTIVE and you my friend are one SPENT fool.:thumbsup::thumbsup:

[GreenLeaf420]

SHANTIBABA OVER HERE IS A:D

[midlifecrisis]

OK, then let's get this straight, you can determine sex after germination but before leaves appear? at what cost per plant, do I have to send my plants to you? or just a small tissue sample?
If I need to understand all that science talk and have expensive equipment, then I surely can't do this myself......is it like a pregnancy test kit???

I guess I am not sure what one would be paying for?

[thegenemaster]

It takes me about 4-6 hours to isolate the DNA from 96 plants. Next day I set up the reactions. I have them set up as a master mix. I add 1ul of isolated DNA to each reaction and add 39up of my master mix that I have determined experimentally will work every time. I can do 96 reactions at once in my thermocycler. Run them out on 3-4 gels for an hour. Take a photo and I'm done. Ten dollars a plant is a very reasonable rate and at that rate I make just under $1000 for less than a days work. I have an abundance of enzyme and my specific choice of forward and reverse primers so I can do literally thousands of reactions. I only have to pay for pipet tips and microfuge tubes. Both of which are cheap if you buy them in bulk.

[thegenemaster]

of the embryo itself. But why would you want to sacrifice the embryo. I wait until after the first set of true leaves appear so that the plant can sustain itself. Then I take both cotyledons, the "first leaves" The lobular shaped leaves that come out of the embryo.

[GreenLeaf420]

It takes me about 4-6 hours to isolate the DNA from 96 plants. Next day I set up the reactions. I have them set up as a master mix. I add 1ul of isolated DNA to each reaction and add 39up of my master mix that I have determined experimentally will work every time. I can do 96 reactions at once in my thermocycler. Run them out on 3-4 gels for an hour. Take a photo and I'm done. Ten dollars a plant is a very reasonable rate and at that rate I make just under $1000 for less than a days work. I have an abundance of enzyme and my specific choice of forward and reverse primers so I can do literally thousands of reactions. I only have to pay for pipet tips and microfuge tubes. Both of which are cheap if you buy them in bulk.

WOW SHANTIBABA you should really read the rules you are looking to get banned. Nice fist impression:thumbsup:

[midlifecrisis]

It takes me about 4-6 hours to isolate the DNA from 96 plants. Next day I set up the reactions. I have them set up as a master mix. I add 1ul of isolated DNA to each reaction and add 39up of my master mix that I have determined experimentally will work every time. I can do 96 reactions at once in my thermocycler. Run them out on 3-4 gels for an hour. Take a photo and I'm done. Ten dollars a plant is a very reasonable rate and at that rate I make just under $1000 for less than a days work. I have an abundance of enzyme and my specific choice of forward and reverse primers so I can do literally thousands of reactions. I only have to pay for pipet tips and microfuge tubes. Both of which are cheap if you buy them in bulk.



OK, I see, cut off your cotyledons and send them in a self addressed, stamped envelope along with your ten dollars and you will tell me the sex of my plant? Is that it?

[thegenemaster]

I have a background in biology, botany, cell signal transduction and biochemistry. I went to school for along time and I have worked in the lab for even longer. Yes I expect to make a few bucks on helping others thin the males from their herds quickly and efficiently.

It requires a very small sample of tissue to do the analysis. I'm really just trying to figure this whole thing out. I know I can do it I have done it many times. Now I'm trying to find a way to help others with the same dilemma I had... how do I make sure I don't waste time space energy and resources on a plant I am just gonna pitch. In my case it was specifially space. Nothing worse than a male occupying valuable grow space. At this point I have gone to my local hydro store where the guy there was very geeked about this and encouraged me to let others know I could do it. So now thats what I'm doing.

[thegenemaster]

hydro store is. I'll go by there and give him my cards and sample bags and you can go in there and pick up both. Call me e-mail me and arrange to get your samples to me. As I said I just starting trying to figure out how to do this. First few guys I did analysis for I actually went and picked up their samples and sent them the results as a picture and message letting them know which plants were males. This also apparently will identify hermaphrodites.

[midlifecrisis]

"Hey look I'm a scientist not business person"



I will not disagree with any of the science aspect, but the business model defiantly needs some work..........I would suggest working that out because if you really believe in this idea then by all means make some money at it.......sort of like cultivating a message board because you believe in it and then making some money on businesses that would like to advertise to your members.......:jointsmile:

[GreenLeaf420]

If this is a sales post, it's in violation if site rules. Please read the forum guidelines concernig unauthorized advertising, and contact the site admin if you would like to advertise legitimately, thanks! :)

Sex in cannabis is 'set' during the very early seedling stage. You can easily force 99.9%+ female by simply pollinating a 'normal' female with pollen from a female that has, either naturally or through photoperiod manipulation or even chemical treatment, produced male sex parts. These 'nanners', as growers call them, are interesting in that, while they produce pollen that by all appearances is in fact viable pollen, it is physically impossible for it to contain a Y chromosome. Therefore, all offspring will be female.

Google 'feminizing seeds' and you will eventually find an interesting article by the breeder Soma on this subject.

STINK FIX THIS GUY!!!!

[thegenemaster]

I'm really not trying to get rich. I developed this for myself. I would have done it even if no one else wanted to pay for me to do it. If I can make a few bucks to help pay off credit cards that would be great.

Just as importantly I have a 20yr old son who is studying to be biochemist as well. I'd love to give him some experience so I have taught him to do the analysis. He knew the principles for class and actually had even isolated plant DNA before. I want to give him experience and let him put it on his resume saying he worked for Green Molecular Solutions(thats me) genotyping plants to help farmers facilitate breeding. Of course he'll get payed decently for doing it too.

So I hope that helps you better understand my business model, or lack there of.

[GreenLeaf420]

Pal you need a infomercial you are really starting to get on my nerves.....

Maybe you did not understand all the warnings from others and myself.

Like I said you should read the rules and re-think what you are saying.

You don't fit the type so I question your integrity greatly you freek me out.. I'm done commenting the MODS will just ban you when they come back.....

[stinkyattic]

I guarantee it works. I am set up to analyze a lot of plants and I can do it for a very reasonable price. Please give me some feed back on this. Thanks!
^First sales offer

If this is a sales post, it's in violation if site rules. Please read the forum guidelines concernig unauthorized advertising, and contact the site admin if you would like to advertise legitimately, thanks!
^First warning

alot of people that would benefit form this service and would be happy to pay a reasonable fee for the service. Obviously I'm not going to do it for free.
^Second sales offer

I'm not trying to get rich.. and yes I have a fucking day job. Man I have just come this form this morning and I have apparently hit the asshole motherload.
^Oh hey be nice now
I'm only here to lend my expertise to fellow growers if they so desire it. Sorry I can't do it for free but I'm not going to go broke helping others out. .
^Third sales offer

you can go in there and pick up both. Call me e-mail me and arrange to get your samples to me.
4th sales offer, also breaking the big 'email me' no no...
I see our time together has come to an end.