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Team:University of Alberta/Plant Project
From 2008.igem.org
(→IGEM's First Binary Vector) |
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[[Selection of Primary Transformants]]<br> | [[Selection of Primary Transformants]]<br> | ||
[[Transformation Protocol]] | [[Transformation Protocol]] | ||
| + | |||
| + | ==Time line== | ||
| + | A new tray of plants will be planted every week. | ||
| + | |||
| + | '''Week 1'''(Before Biobricks Arrive)<br> | ||
| + | PCR our the resistance gene and promoter from the Pcambia vector<br> | ||
| + | Use Site directed mutagenesis to remove NOTI sites from the vector<br> | ||
| + | |||
| + | '''Week 2'''(biobricks have arrived)<br> | ||
| + | Digest new biobrick and confirm it is what it is.<br> | ||
| + | Insert Plant biobrick into the pCambia vector that has had the NOTI sites removed<br> | ||
| + | Transform Vector into Agrobacterium<br> | ||
| + | Confirm Transformation<br> | ||
| + | Create bulk culture of Agrobacterium for plant transformation | ||
| + | |||
| + | '''Week 3''' | ||
| + | Transform Plants ( That are 3 weeks old) | ||
| + | Make another bulk culture for next weeks transformation | ||
| + | '''Week 4-6''' | ||
| + | Keep transforming plants on tray every week to increase chances. | ||
| + | Harvest seeds of transformed plants and replant them.(continue as seeds come into maturity) | ||
| + | '''Week 6-End''' | ||
| + | select for homozygous plants | ||
Revision as of 16:23, 20 June 2008
Contents |
IGEM's First Binary Vector
Our Binary Vector is a modified pCambia 0380 binary vector [link] We have added in our own poly linker which includes the proper prefix, suffix and primers to coincide with the IGEM regulations. We also added a CAM resistance cassette.
The First Plant Project Ever Submitted to IGEM
We hope to demonstrate that using biobricks in plants is not only useful but feasible in the restraints of this competition. We also hope to use the tools that we make to insert our toxin detection vector into plants.
Plant Protocol
Growing Plants
Selection of Primary Transformants
Transformation Protocol
Time line
A new tray of plants will be planted every week.
Week 1(Before Biobricks Arrive)
PCR our the resistance gene and promoter from the Pcambia vector
Use Site directed mutagenesis to remove NOTI sites from the vector
Week 2(biobricks have arrived)
Digest new biobrick and confirm it is what it is.
Insert Plant biobrick into the pCambia vector that has had the NOTI sites removed
Transform Vector into Agrobacterium
Confirm Transformation
Create bulk culture of Agrobacterium for plant transformation
Week 3 Transform Plants ( That are 3 weeks old) Make another bulk culture for next weeks transformation Week 4-6 Keep transforming plants on tray every week to increase chances. Harvest seeds of transformed plants and replant them.(continue as seeds come into maturity) Week 6-End select for homozygous plants
