http://bioinformatics.dnalc.org/gmo/animation/pdf/Detecting%20GM%20Foods%20by%20PCR.pdf
I use this protocol for extracting genomic DNA from plants daily. Its not ultrapure but will last a while. I've had genomic arabidopsis DNA using this protocol for three years and its still useable.
Then design primers to isolate the gene along with restriction digest sites (if you are doing traditional cloning), run a pcr, cut and clean the bands, digest it along with the plasmid, run another gel to purify the sample, ligate it together, transform into bacteria.
If you want to transfer genes u need to find some agrobacterium from a lab and a transformation vector. Then clone in the gene along with a promoter (CaMV or zea mays ubiquitin for example). You should also try your hand at tissue culture and start with a simple organism. Tobacco is my main plant. Don't be fooled by the simplicity of arabisopsis floral dip. Its terribly inefficient and the flowers are tiny and do not last long. See attached images for tobacco flower. It takes five weeks to get a GMO via tissue culture post agro transformation. I do this stuff all the time. If you want more info, feel free to ask. I love helping people learn more about plant genetic engineering. Best of luck and don't forget to have fun!
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
I use this protocol for extracting genomic DNA from plants daily. Its not ultrapure but will last a while. I've had genomic arabidopsis DNA using this protocol for three years and its still useable.
Then design primers to isolate the gene along with restriction digest sites (if you are doing traditional cloning), run a pcr, cut and clean the bands, digest it along with the plasmid, run another gel to purify the sample, ligate it together, transform into bacteria.
If you want to transfer genes u need to find some agrobacterium from a lab and a transformation vector. Then clone in the gene along with a promoter (CaMV or zea mays ubiquitin for example). You should also try your hand at tissue culture and start with a simple organism. Tobacco is my main plant. Don't be fooled by the simplicity of arabisopsis floral dip. Its terribly inefficient and the flowers are tiny and do not last long. See attached images for tobacco flower. It takes five weeks to get a GMO via tissue culture post agro transformation. I do this stuff all the time. If you want more info, feel free to ask. I love helping people learn more about plant genetic engineering. Best of luck and don't forget to have fun!
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
From: Nico B.
Sent: 11/23/2014 7:03 PM
To: diybio@googlegroups.com
Subject: [DIYbio] Isolating sequences from scratch
Hi,
I'm maybe 4 months into any semblance of synbio/molecular bio/bio hacking and have a few projects running because of the amazing support at Berkeley Biolabs and this online community, so for that thanks.
Maybe you've seen pictures of the Diphylleia grayi (skeleton flower) that have been making their way across the internet. I would love to learn to isolate genes -in this instance, that which is responsible for the effect of translucency of the pedals when it comes into contact with water- and have no idea as to how to do this once I've got the physical plant in my possession.
HALP!
Thanks,
Nico
-- I'm maybe 4 months into any semblance of synbio/molecular bio/bio hacking and have a few projects running because of the amazing support at Berkeley Biolabs and this online community, so for that thanks.
Maybe you've seen pictures of the Diphylleia grayi (skeleton flower) that have been making their way across the internet. I would love to learn to isolate genes -in this instance, that which is responsible for the effect of translucency of the pedals when it comes into contact with water- and have no idea as to how to do this once I've got the physical plant in my possession.
HALP!
Thanks,
Nico
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