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OriCiro Cell-Free Cloning System

®

Large Circular DNA Construction Tool

01

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.

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02

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.

Here's a video explaining the principles and excellent features of OriCiro Cell Free Cloning System

PRODUCT OVERVIEW

OriCiro® Assembly Kit

01

REACTION

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.

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FEATURES

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Efficient ligation
by isothermal
enzymatic reactions.

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Up to 50 DNA fragments
can be assembled together
as designed.

03

DISTINCTIVE ADVANTAGES

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Up to
50 kb DNA fragment
can be assembled.

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.

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Assembly of 0.6 kb × 10 fragments

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otal 6 kb,
Each fragments have 40 bp overlapping ends

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.

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02

FEATURES

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in vitro reconstructionof the E. coli genomereplication system.

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Up to 10,000 times
more accurate amplification
than PCR.

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More than 1 billion times
amplification from 1 circular
DNA molecule by isothermal
reaction for several hours.

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Consecutive amplification
for many generation (passage)
is possible by subculturing
in a new reaction solution.

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Amplification of large circular
DNA exceeding 1 Mbp.
※Our products are optimized for up to
50 kb.

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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.

03

DISTINCTIVE
ADVANTAGES

Please refer to the table below for the advantages of OriCiro amplification technology over the conventional methods of E. coli cloning and PCR.

※1 Technology under development to mach E. coli cloning.