Editing Gibson Assembly (section)

== Gibson Method, aka VLIC, aka one pot isothermal assembly ==

Original protocol from: [http://www.nature.com/nmeth/journal/v6/n5/full/nmeth.1318.html this paper], doi:10.1038/nmeth.1318

#PCR up your fragment of choice and gel purify
#Thaw a 15 μl assembly mixture aliquot and keep on ice until ready to be used.
#Add 5 μl of DNA to be assembled to the master mixture. The DNA should be in equimolar amounts. Use 10-100 ng of each ~6 kb DNA fragment. For larger DNA segments, increasingly proportionate amounts of DNA should be added (e.g. 250 ng of each 150 kb DNA segment).
#Incubate at 50 °C for 15 to 60 min (60 min is optimal).
#Transform as usual