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From 2011.igem.org

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Revision as of 09:28, 12 July 2011

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 :)