Revision as of 09:32, 12 July 2011

Gibson assembly

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 :)

Revision as of 09:31, 12 July 2011 (view source) Nguenat (Talk \| contribs) ← Older edit		Revision as of 09:32, 12 July 2011 (view source) Nguenat (Talk \| contribs) Newer edit →
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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)