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Team:Washington/Magnetosomes/GibsonVectors
From 2011.igem.org
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==What happened last year?== | ==What happened last year?== | ||
| - | The Gibson Cloning method is definitely not a new terminology to the iGEM community. This cloning method | + | The Gibson Cloning method is definitely not a new terminology to the iGEM community. This cloning method was published in Nature protocols 2009 and it has gained a lot of attention from the iGEM community since then. The main reason behind is that it allows multiple inserts in one time isothermal reaction during the cloning process, which does not only saves a lot of time and efforts, but also allows more complexity when building the circuit. |
| - | + | In 2010, the Cambridge iGEM team proposed the proposed a BioBrick standard for Gibson Assembly (RFC 57) which enables simultaneous assembly of multiple fragments with no scar sequences. We found that this Biobrick Standard was incapable of giving high yields even in two-fragments assembly. The primary problem is the self-complementarity of the two NotI sequences embedded in the BioBrick prefix and suffix which prevents the insert from being incorporated into the vector. | |
| + | [[File:Igem2011 biobrick NotI.png|400px|middle]] | ||
| + | |||
| + | Therefore, to optimize the method, a “gibson reaction compatible” prefix and suffix were also designed based on BglBrick standard to increase the cloning efficiency. | ||
== What about this year? == | == What about this year? == | ||
Revision as of 21:24, 22 September 2011
Gibson Vector Toolkit
About Gibson Cloning/Assembly
This is a one-step isothermal reaction method for assembling overlapping DNA fragments. Please see Enzymatic assembly of DNA molecules up to several hundred kilobases, Gibson et al. (2009). Outline; What is gibson (and diagram from the slide)
New vectors... a diagram a table showing the antibiotics, copy number
This is an outline!!
What happened last year?
The Gibson Cloning method is definitely not a new terminology to the iGEM community. This cloning method was published in Nature protocols 2009 and it has gained a lot of attention from the iGEM community since then. The main reason behind is that it allows multiple inserts in one time isothermal reaction during the cloning process, which does not only saves a lot of time and efforts, but also allows more complexity when building the circuit.
In 2010, the Cambridge iGEM team proposed the proposed a BioBrick standard for Gibson Assembly (RFC 57) which enables simultaneous assembly of multiple fragments with no scar sequences. We found that this Biobrick Standard was incapable of giving high yields even in two-fragments assembly. The primary problem is the self-complementarity of the two NotI sequences embedded in the BioBrick prefix and suffix which prevents the insert from being incorporated into the vector.
Therefore, to optimize the method, a “gibson reaction compatible” prefix and suffix were also designed based on BglBrick standard to increase the cloning efficiency.
What about this year?
Seeing that this is such a great method to do cloning...we CONTINUE WITH THE INVESTIGATION! AND WE MADE/SUBMITTED MORE VECTORS that are compatible with Gibson cloning in a hope to ease everyone’s pain when doing cloning. (bring the iGEM to a differnt level of cloning)
Next level of iGEM project: more complex circuit.
And we decided to look at magnetosome
