OriCiro Cell-Free Cloning system
Assemble to Amplify up to 50kb DNA in a day

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.

APPLICATION

• 01Artificially designed viral vector and plasmid construction

• 02Amplification of long DNA and sequences difficult to amplify by PCR

• 03Alternative to time-consuming and laborious E. coli cloning

• 04Recombinant phage production

• 05Efficient cloning of any DNA sequence, including repeat, cytotoxic and GC-rich sequences

• 06The amplification products can be used directly for E. coli transformation, etc.

PRODUCT OVERVIEW

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.

FEATURES

• 01Efficient ligation by isothermal enzymatic reactions.

• 02Up to 50 DNA fragments can be assembled together as designed.

• 03Up to 50 kb DNA fragment can be assembled.

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OriCiro® Amp Kit

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.

FEATURES

• 01in vitro reconstruction of the E. coli genome replication system.

• 02More than 1 billion times amplification from 1 circular DNA molecule by isothermal reaction for several hours.

  
  

• 03Amplification of large circular DNA exceeding 1 Mbp.※Our products are optimized for up to 50 kb.

• 04Up to 10,000 times more accurate amplification than PCR.

• 05Consecutive amplification for many generation (passage) is possible by subculturing in a new reaction solution.

• 06Amplification product can be transformed into E. coli without purification step.

• 07Sequences with GC-rich which are difficult to amplify by PCR or E. coli can be amplified.
  

OriCiro® Cell-Free Cloning System

Academia/non-profit Customer

Corporate customer