Difference between revisions of "Gibson Assembly"
From GcatWiki
| Line 16: | Line 16: | ||
| − | Insert Size | + | Insert Size Insert:Vector |
| − | 25-300 bp | + | 25-300 bp 5:1 |
| − | 300-1000 bp | + | 300-1000 bp 3:1 |
| − | 1000-3000 bp | + | 1000-3000 bp 1:1 |
Insert Calculation | Insert Calculation | ||
Revision as of 16:44, 12 July 2012
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
