Gibson Assembly
From GcatWiki
Assembly Recipe and Methods
2ul NEB Gibson MM _ul Vector (Always 20ng) _ul Insert (Calculate insert amount below) _ul dH2O
4ul Total Volume
Incubate at 50 degrees Celcius for approximately 1 hour.
Transform 1 ul w/ 40ul Cells diluted to no more that 8 fold
Insert Size Insert:Vector
25-300 bp 5:1
300-1000 bp 3:1
1000-3000 bp 1:1
Insert Calculation
Insert (ng)=(ng of vector) (Insert bp/Vector bp)(1 or 3 or 5) e.g. (20)(75/2900)(5)= 2.6 ng