"

From 2013.igem.org

(18.10)

HM17-FS_78

Annealing temps: 62°C, 60°C, 57°C, 55°C
Elongation time: 2:10

Screening remaining Gibson electroporated

60 colonies were picked and screened with primers FS_73 and FS_95 (annealing 66 touchdown, 62)

Gibson assembly

Reagent	Volume added (µl)
BB (FS..)	0.61
DN11-FS66	1.00
FS_67-FS_68	1.35
FS_69-FS_70	6.34
FS_71-HM08	0.70
Gibson Master Mix 2x	10