Welcome to OriCiro Genomics
A pioneer in cell-free synthesis and amplification of genome-scale large DNA for synthetic biology
Enzymatic DNA synthesis on demand
In-vitro reconstitution of chromosome replication cycle of E. coli
Assemble to Amplify
50 DNA fragments,Total 50 kb
in hours
OriCiro® Cell-free Cloning system
Enzymatic DNA Synthesis Technology
DISTINCTIVE ADVANTAGES
APPLICATION
・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 repeat, cytotoxic and GC-rich sequences
・The amplification products can be used directly for E. coli transformation, etc.
TECHNOLOGY
OriCiro® 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.
OriCiro® Amp Kit
01
REACTION
OriCiro® Amp Kit contains 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 1 μg circular DNA can be yielded as supercoiled form per 10 μl reaction.
For more detailed explanation, refer to the relevant publication.
02
FEATURES
in vitro reconstructionof the E. coli genomereplication system.
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.
Consecutive amplification
for many generation (passage)
is possible by subculturing
in a new reaction solution.
Amplification of large circular
DNA exceeding 1 Mbp.
※Our products are optimized for up to
50 kb.
Amplification product
can be transformed
into E. coli without
purification step.
Sequences with GC-rich which are difficult to amplify by PCR or
E. coli can be amplified.
Application Note
OriCiro® Assembly Kit
In OriCiro® Assembly Kit, DNA fragments are assembled via overlapping ends having homologous sequences. First, each end of the fragments is digested by exonucleases to expose a single stranded homologous sequence. Then, corresponding fragments having the homologous sequence each other are annealed by enzymes. The kit assembles up to 50 DNA fragments simultaneously in a single isothermal enzymatic reaction. The assembly products are subjected directly to OriCiro® Amp Kit for selective amplification of your target circular DNA.
01
REACTION
02
FEATURES
03
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.
DISTINCTIVE ADVANTAGES
Ten fragments of 0.6 kb having 40 bp overlapping ends were assembled using OriCiro Assembly kit. The OriCiro kit (OriCiro) showed an assembly product of all ten fragments. DNA fragments before (-) or after (+) the assembly reaction were analyzed in agarose gel electrophoresis.
Assembly of 0.6 kb × 10 fragments
