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Team:Alberta/Achievements/Overview
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| - | The octamer demonstrated that we could create long constructs out of pieces of DNA, so the next step was to construct a fully functional plasmid and test drive it in a cell. To do this, we started with a short construct that contained all the required parts of a plasmid. This also allowed us to test out the parts we had made specifically for this purpose. | + | The octamer demonstrated that we could create long constructs out of pieces of DNA, so the next step was to construct a fully functional plasmid and test drive it in a cell. [[Image:Alberta Tformplasmid.jpg|right|250px]] To do this, we started with a short construct that contained all the required parts of a plasmid. This also allowed us to test out the parts we had made specifically for this purpose. |
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This plasmid is seen on the right and contains everything needed to function in vivo. | This plasmid is seen on the right and contains everything needed to function in vivo. | ||
Revision as of 03:30, 27 October 2010
BioBytes 2.0
The first major accomplishment of our project was to optimize the BioBytes 2.0 standard for parts and the assembly of those parts into useful transformable constructs. This was done very early on, but was essential for the progression and success of the remainder of the project.
DNA Attachment
The next major step in the development of GENOMIKON was to successfully attach the BioBytes 2.0 anchor to poly-T iron micro beads. This was demonstrated by attaching a 1kbp anchor-AB KanR to the iron micro beads, then eluting it off of the beads.
As we can see, all of the 1kbp anchor-AB KanR is removed in the initial elution and none remains attached to the beads in the subsequent wash step. This is also true of the two BioBytes 2.0 construct seen on the right that is a 2kbp piece added to a 1kbp piece.
Octamer Construction
Our next major accomplishment was the assembly of a 12kbp construct made of 8 pieces of DNA using the BioBytes 2.0 assembly method. The octamer was assembled on an iron micro bead, starting with the anchor, and adding alternating 1kbp pieces and 2kbp pieces until the full 12kbp construct was achieved. This is seen in the third lane. A tetramer of the same construction is seen in the second lane.
It can be observed that the the major product of both the octamer and tetramer constructs was the complete construct. This was an exciting gel, as the octamer was constructed in under 3 hours - a feat that would have taken more than an entire summer using the BioBrick method.
Transformation of E. coli with a Constructed Plasmid
The octamer demonstrated that we could create long constructs out of pieces of DNA, so the next step was to construct a fully functional plasmid and test drive it in a cell.
This plasmid is seen on the right and contains everything needed to function in vivo.
High School Student Transformations
Once we had transformed E. coli with constructs we made using BioBytes 2.0, we wanted to test the educational value of the project. To do this, we completed the same experiment with five high school students using precision droppers.
