Some of the genes have been sequenced and the others have known chemical structures which allow the amino acid sequence to be determined... and that allows you to work back thru the mRNA that would create those amino acid chains and then from there work back to the DNA which would create the mRNA.

Gene Databases

Figuring out the DNA sequence from an amino acid chain creates several possible sequences, but with degenerate primers (primer nuecleotides that can bond to more than 1 DNA nuecleotide types), you can use 1 primer set and isolate a gene without knowing its complete sequence.

I've never grafted. Because DNA doesn't transfer to cuttings/seedlings via grafting, it really doesn't have much value to me...

I've been too busy to post much of the latest work, but I'll get around to it. I lost a bunch of pictures and video because I thought they were older footage and I deleted them by mistake. I'm going to redo the whole DNA extraction, isolation and transfection... I'm going to make my own gel electrophorese holder that has a removable bottom, so I'll be able to pop the whole gel out... And I want to do a time lapse video of a gel run.

I've gotten good results so far and have 2 batches of cell colonies that tested positive for 3 of the 4 DNA transfected sequences. Those cells will be grown out and used for the next round of transfection. Its possible that they already carry enough genes to create THC. I'll probably have the new video done before they've grown out enough to tell, but the promoter should be kicking in the genes in every part of the plant, so even leaves should get you high...

-Loki

What kind of nutrient based medium are you growing these altered cells in?


What method did you use for transfection?

Sorry for all the questions I'm just really interested and curios.

The reason I asked about grafting is because I'm actually attempting grafting experiments right now in a green house under the supervision of of my boss. If these experiment work out my next step will be to attempt them with cannabis.

The medium is a Murashige and Skoog mix with vitamins and ampicillin.

I tried a few transfection methods, but the one that actually worked was TurboFect. It's kind of expensive, but you can find guys selling Fermentas products on eBay and some will sell half or a quarter of a kit for $75-100. Each transfection takes just a single drop of the transfection reagent, so a little goes a long way.

-Loki

Loki I am honestly glued to this thread bro. I hope you don't lose interest in this. I really wanna see the outcome.

Very impressive I am also glued to this thread.

There'll be more to come soon... well, maybe a month tops...

I bought my own oligo synthesizer, so I can make my own primers and even artifical genes. It's pretty fast for an older 8 column unit. Cost just under a grand, delivered, but it's wicked cool to be able to crank out my own DNA sequences... gives me a techno-chubby...

-Loki

LOL a techno-chubby? Thats a new one.

Damn, interesting stuff!! You should try makeing some super strains man this is cool and all but reminds me why I dropped Biology, lol Have Fun Bro!!!

Peace -

Loki Amazing,
I am giving you a standing ovation, if the government won't do research Loki will. I just got done reading the whole thread and I love how you are educating others on the process, might be a slightly "Dumbed Down" version but still I have really enjoyed your data and info keep up the good work.

Thanks, it helps me make sure I've understood things to then try and explain them.

Now that I can make my own primers, things will be so much easier. Once it's done, I'm going to post all the primers used to isolate the target genes and the protocols for isolating, extracting and transfecting them. That sounds pretty impressive, but it's actually not as complicated as most people think. Anyone with a high school education and a couple grand will be able to reproduce all my work. I'll send out sample plants to people who can't or don't want to do it themselves. Once people see THC in a non-marijuana plant, some will want to put THC in their favorite plant(s) and I'll show them how...

-Loki