Assembly Related Protocols
From 2008.igem.org
(Difference between revisions)
(New page: '''Newest EcoRI/BamHI Transfer method (7/31/08)''' Digestion: 0.5ul plasmid in entry vector 0.5ul EcoRI 0.5ul BamHI 1ul NEB2 + ATP buffer 7.5ul of water incubate the digestion for 1hr....) |
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Digestion: | Digestion: | ||
- | 0.5ul plasmid in entry vector | + | *0.5ul plasmid in entry vector |
- | 0.5ul EcoRI | + | *0.5ul EcoRI |
- | 0.5ul BamHI | + | *0.5ul BamHI |
- | 1ul NEB2 + ATP buffer | + | *1ul NEB2 + ATP buffer |
- | 7.5ul of water | + | *7.5ul of water |
- | incubate the digestion for 1hr. in the thermocycler, and 20 min for heat kill. | + | *incubate the digestion for 1hr. in the thermocycler, and 20 min for heat kill. |
No purification step. | No purification step. | ||
Ligation: | Ligation: | ||
- | 1ul of dilute assembly vector (or 0.5ul if undiluted) | + | *1ul of dilute assembly vector (or 0.5ul if undiluted) |
- | 0.5ul ligase | + | *0.5ul ligase |
Transform. (same as usual) | Transform. (same as usual) |
Revision as of 03:03, 2 August 2008
Newest EcoRI/BamHI Transfer method (7/31/08)
Digestion:
- 0.5ul plasmid in entry vector
- 0.5ul EcoRI
- 0.5ul BamHI
- 1ul NEB2 + ATP buffer
- 7.5ul of water
- incubate the digestion for 1hr. in the thermocycler, and 20 min for heat kill.
No purification step.
Ligation:
- 1ul of dilute assembly vector (or 0.5ul if undiluted)
- 0.5ul ligase
Transform. (same as usual)