Difference between revisions of "Gibson Assembly"
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(Created page with '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 degre…') |
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2ul NEB Gibson MM | 2ul NEB Gibson MM | ||
| + | |||
_ul Vector (Always 20ng) | _ul Vector (Always 20ng) | ||
| + | |||
_ul Insert (Calculate insert amount below) | _ul Insert (Calculate insert amount below) | ||
| + | |||
_ul dH2O | _ul dH2O | ||
| Line 10: | Line 13: | ||
Incubate at 50 degrees Celcius for approximately 1 hour. | Incubate at 50 degrees Celcius for approximately 1 hour. | ||
| − | Transform | + | Transform 1ul w/ 40ul Cells diluted to no more that 8 fold |
| − | Insert Size | + | Insert Size; Insert:Vector |
| − | 25-300 bp | + | |
| − | 300-1000 bp | + | 25-300 bp; 5:1 |
| − | 1000-3000 bp | + | |
| + | 300-1000 bp; 3:1 | ||
| + | |||
| + | 1000-3000 bp; 1:1 | ||
Insert Calculation | Insert Calculation | ||
Insert (ng)=(ng of vector) (Insert bp/Vector bp)(1 or 3 or 5) | Insert (ng)=(ng of vector) (Insert bp/Vector bp)(1 or 3 or 5) | ||
| + | |||
e.g. (20)(75/2900)(5)= 2.6 ng | e.g. (20)(75/2900)(5)= 2.6 ng | ||
Latest 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
