Engineering of customized meganucleases via in vitro compartmentalization and in cellulo optimization

. Author manuscript; available in PMC: 2015 May 1.

Published in final edited form as: Methods Mol Biol. 2015;1239:105–132. doi: 10.1007/978-1-4939-1862-1_6

2 × Gibson Assembly Master Mix	5 μl
PCR fragment encoding variant endonucleases	30-40 ng
Linearized pEndo plasmid	60-75 ng
Water	Added to make a final volume of 10 μl