Results 1 to 10 of 24
Threaded View
-
06-14-2008, 02:38 AM #6OPJunior Member
Using genetics to determine a plants sex
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.
Similar Threads
-
Seed Demonstration - Good Genetics vs Bad Genetics
By Minnesnowta in forum Basic GrowingReplies: 0Last Post: 01-17-2012, 02:43 AM -
How do I determine my first sample of the day?
By itsonnowgirl in forum Drug TestingReplies: 2Last Post: 01-23-2010, 07:12 AM -
How do I determine light distance from plants.
By latewood in forum Grow FAQsReplies: 0Last Post: 02-15-2007, 08:26 AM -
How to determine what u have
By Booshkie in forum Marijuana MethodsReplies: 8Last Post: 01-16-2007, 09:39 AM -
Please help me determine the sex of both plants
By Sidd in forum Basic GrowingReplies: 8Last Post: 09-02-2004, 09:02 AM