pET-43.1 Ek/LIC: Difference between revisions
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==At a Glance {{HelpIcon|Help:Plasmid}}== | ==At a Glance {{HelpIcon|Help:Plasmid}}== | ||
[[Image:#PET43.1EkLIC.jpg|thumb|right|300px|Image provided by [http://www.emdbiosciences.com/docs/docs/PROT/TB318.pdf Novagen]]] | |||
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==Notes== | ==Notes== | ||
==History== | ==History== | ||
Revision as of 22:49, 26 November 2007
At a Glance
[[Image:#PET43.1EkLIC.jpg|thumb|right|300px|Image provided by Novagen]]
| Type |
|---|
| Origin of Replication |
| Copy # |
| Markers |
| Link to Sequence |
| Comments |
| edit table |
Notes
History
Ligation-independent cloning (LIC) was developed for directional cloning of PCR products without restriction enzyme digestion or ligation reactions[1][2]. LIC takes advantage of the 3’→5’ exonuclease activity of T4 DNA polymerase to create very specific 12 to 15 nucleotide single-stranded overhangs in the vector and the insert, so that the vast majority of annealed products consist of the desired molecules. The annealed LIC vector and insert are transformed into competent E. coli cells, and covalent bonds are formed at the vector-insert junctions within the cell to yield circular plasmid. Directional cloning of the insert is achieved with minimal non-recombinant background, and cloning is so efficient that virtually all of the resulting colonies contain the desired recombinant.
Sources
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References
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Sequence
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