Using genetics to determine a plants sex

[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

[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: