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OriCiro Cell-Free Cloning System
This kit includes 5 reactions of OriCiroTM Assembly and 10 reactions of OriCiroTM Amp.
Contents & storage
OriCiroTM Cell-Free Cloning System is an innovative tool enabling cell-free assembly and amplification of circular DNA molecules without E. coli transformation and culture. This product can not only streamline your workflow dramatically but also widen the scope of your genetic engineering techniques. This product is composed of OriCiro Assembly Kit and OriCiro Amp Kit and designed to produce the maximum effect when used in combination although each kit can be used independently.
Artificially designed viral vector and plasmid construction
Amplification of long DNA and sequences difficult to amplify by PCR
Alternative to time-consuming and laborious E. coli cloning
Recombinant phage production
Efficient cloning of any DNA sequence, including cytotoxic and GC-rich
DNA fragments are assembled via overlapping ends having homologous sequences. First, each end of the fragments is digested by exonuclease to expose a single stranded homologous sequence. Then, corresponding fragments having the homologous sequence each other are annealed by enzymes.
Efficient ligation by isothermal enzymatic reactions.
Up to 50 DNA fragments can be assembled together as designed.
Up to 50 kb DNA fragment can be assembled
26 kinds of purified enzymes that are essential for the E. coli chromosome replication cycle.
This Kit enables to amplify exponentially circular DNA having oriC (E. coli chromosomal origin) in an isothermal condition even from a single template molecule. Up to 1ug circular DNA can be yielded as supercoiled form per 10 ul reaction.
More than 1 billion times amplification from 1 circular DNA molecule by isothermal reaction for several hours
Up to 10,000 times more accurate amplification than PCR (Taq polymerase)
Sequences with GC-rich or cytotoxic which are difficult to amplify by PCR or E. coli can be amplified.
1. T. Mukai, T. Yoneji, K. Yamada, H. Fujita, S. Nara, M. Su'etsugu, Overcoming the Challenges of Megabase-Sized Plasmid Construction
in Escherichia coli, ACS Synthetic Biology, 2020, 9 (6), 1315-1327
2. M. Su’etsugu, H. Takada, T. Katayama, H. Tsujimoto, Exponential propagation of large circular DNA by reconstitution of a chromosomereplication cycle, Nucleic Acids Research, 2017, 45 (20), 11525–11534
3. T. Hasebe, K. Narita, S. Hidaka, M. Su'etsugu, Efficient Arrangement of the Replication Fork Trap for In Vitro Propagation of
Monomeric Circular DNA in the Chromosome-Replication Cycle Reaction. Life, 2018, 8 (43)