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Team:EPF-Lausanne/Protocols/Gibson assembly
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Protocol adapted from Nikolaus Obholzer (https://wiki.med.harvard.edu/SysBio/Megason/IsothermalAssembly) | Protocol adapted from Nikolaus Obholzer (https://wiki.med.harvard.edu/SysBio/Megason/IsothermalAssembly) | ||
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* Heat at 55°C for 45 minutes (there is a programm written in the iGEM folder) | * Heat at 55°C for 45 minutes (there is a programm written in the iGEM folder) | ||
* Purify with Qiagen PCR purification kit | * Purify with Qiagen PCR purification kit | ||
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| + | Now the samples are ready to be transformed. | ||
PS: if someone has a less complicated method, please change the protocol :) | PS: if someone has a less complicated method, please change the protocol :) | ||
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| + | {{:Team:EPF-Lausanne/Templates/Footer}} | ||
Revision as of 09:31, 12 July 2011
Agarose gel electrophoresis
Protocol adapted from Nikolaus Obholzer (https://wiki.med.harvard.edu/SysBio/Megason/IsothermalAssembly)
- Take the different DNA parts you want to assemble and measure their concentration with Nanodrop
- We need equal molecuar ratios! Divide the concentration measured above [ng/ul] by the length of each part to have equivalent numbers
- Based on these numbers, calculate the ul you need of each to have the same amount of molecules
- Take 5 ul of the mixed solution and add 15 ul of Gibson mastermix
- Heat at 55°C for 45 minutes (there is a programm written in the iGEM folder)
- Purify with Qiagen PCR purification kit
Now the samples are ready to be transformed.
PS: if someone has a less complicated method, please change the protocol :)
