Gibson Assembly

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