Gibson Assembly

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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 1ul 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