The Overlap Cloner DNA Cloning Kit provides a simple and fast directional cloning of one or more DNA fragments into any vectors. The main advantage of Overlap Cloner is that DNA fragments can be precisely cloned as intended without any other additional bases, and several DNA fragments with overlapping sequences can be connected by single reaction.
This system requires no site-specific recombination sites or extra DNA sequences as do in Cre/LoxP or Integrase/exisionase reactions. It eliminates the need for restriction enzymatic and DNA ligation steps in DNA cloning process.The enzyme mix in this kit recognizes more than 15 bp overlapping sequences at each ends of DNA fragment and precisely connects the DNA fragments. This 15 bp overlapping sequences can be generated by PCR amplification with primers.The kit can be used for cloning of up to 6 DNA fragments at the same time. But, the transformation efficiency can be decreased as the number of fragments are increased.
Kit Components
- 100 ㎕ Overlap Cloner Enzyme Mix
- 500 ㎕ Overlap Cloner 10x Reaction Buffer
- 25 ㎕ Control Vector (50 ng/㎕)
- 25 ㎕ Control Insert (LacZ DNA, 50 ng/㎕)
- Manual
QC tests
- The performance of Overlap Cloner DNA Cloning Kit is monitored routinely on lot numbers. The quality of kit is tested with control vector and insert DNA fragments by the criteria of enzymatic performance, transformation efficiency, and molecular analysis of resulted plasmid DNA from several clones.
Storage Condition
- The Overlap Cloner DNA Cloning Kit should be stored at -20℃ or below. The kit can be stored for up to 12 months without showing any reduction in performance and quality.
Applications :
1. Simple cloning (ligation independent)
2. Easy subcloning to destination vectors
3. Highly efficient library construction
4. Gene fusion (advanced than overlapping PCR)
5. No additional sequences in expression study
6. Vector reconstitution study in one tube
7. One pot reaction of in vitro gene synthesis
Features :
1. No use of restriction enzymes & ligase
2. Rapid reaction time : 30~60 min reaction
3. High efficiency : up to 90% when single insert used
4. Multiple ligation : up to 6 DNA fragments in one reaction
5. Exact in-frame cloning : no additional bases or amino acids
