GoldenGate is a method to assemble multiple DNA fragments together. Each fragment is flanked by specific restriction enzyme sites (BsaI in our case) which allows the restriction enzyme to cut at specific sites. The sticky ends at this cut can base pair with other sticky ends in other fragments. Using different sticky end sequences allows us to assemble multiple DNA fragments in a specific order. We used this method to construct our collagen gene as the gene was too long to synthesize as a single chain.
This method is faster and more precise than other assembly methods. It does not require PCR and can create scarless DNA modifications
Other alternate traditional methods could be to use marine collagen as that’s less expensive to extract than mammalian collagen, but this could lead to further complications with reference to rejection. However, ultimately our experiment is a way to make collagen synthesis cheaper. It is more cost effective to grow a field of soybeans that continuously produce human collagen as the plant needs to be transfected once with the gene and further generations will also have the collagen gene.
Great poster! Is there a certain experiment or element you would need to do to produce a higher DNA concentration to meet the required concentration? Or just repeat the original experiment?
curesymposium.org 
Very nice poster! What is the concentration you are aiming for in order to proceed with the experiment?
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I loved your poster! Can you comment more on what exactly GoldenGate Assembly is and how it differs from other methods of assembly?
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GoldenGate is a method to assemble multiple DNA fragments together. Each fragment is flanked by specific restriction enzyme sites (BsaI in our case) which allows the restriction enzyme to cut at specific sites. The sticky ends at this cut can base pair with other sticky ends in other fragments. Using different sticky end sequences allows us to assemble multiple DNA fragments in a specific order. We used this method to construct our collagen gene as the gene was too long to synthesize as a single chain.
This method is faster and more precise than other assembly methods. It does not require PCR and can create scarless DNA modifications
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Great presentation! Is there a way to make traditional collagen synthesis cheaper?
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Other alternate traditional methods could be to use marine collagen as that’s less expensive to extract than mammalian collagen, but this could lead to further complications with reference to rejection. However, ultimately our experiment is a way to make collagen synthesis cheaper. It is more cost effective to grow a field of soybeans that continuously produce human collagen as the plant needs to be transfected once with the gene and further generations will also have the collagen gene.
LikeLike
Great poster! Is there a certain experiment or element you would need to do to produce a higher DNA concentration to meet the required concentration? Or just repeat the original experiment?
LikeLike