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Advancing Life Sciences Technology

ClonePro™ SLIC Enzyme (CPS1001)

CloneProTM SLIC enzyme (CPS1001) in ATCGbio Life Technology Inc. is developed based on the technique of Sequence and Ligation Independent Cloning (SLIC).  In 20 min reaction, PCR product can be inserted in immediately.
SLIC (Sequence and Ligation Independent Cloning) technique employs in vitro directional homologous recombination at cohesive ends created by 5’→3’ or 3’→5’ exonuclease activity reaction.  It has been developed for years since the first report using T4 DNA polymerase in 1993(1), and this protocol was restated by Mamie Z Li and Stephen J Elledge(2) in 2007. Besides using T4 DNA polymerase, the protocol using exonuclease III(3), T7 exonuclease (4), T5 exonuclease(5) and vaccinia virus DNA polymerase(6) were also reported.  Commercial products are also available using these enzymes and others in proprietary combination.  In-Fusion® (Takara), GENEART® Seamless cloning and assembly enzyme mix (Invitrogen), Gibson Assembly® Master Mix (NEB) are such enzyme kits and are made to optimize exonuclease activity and filled-in polymerase activity. However, those enzymes are more suitable for long homologous (more than 20-30 bp) and long fragment (more than 100 bp).
CloneProTM SLIC enzyme has similar or more effective function as In-Fusion® (Takara), GENEART® Seamless cloning, assembly enzyme mix (Invitrogen), Gibson Assembly® Master Mix (NEB) and others.
                                  5’-exonuclease activity    ------------->      
5’-NNNNNNNNNNNNNNN                                                             NNN-3’     
3’-NN                                                         NNNNNNNNNNNNNNNNNN-5’
                  <---------- 5’-exonuclease activity

 NNNNNNNNNNNNN and NNNNNNNNNNNNNNN are cohesive ends that can anneal spontaneously.

Features of CloneProTM SLIC enzyme
CloneProTM SLIC enzyme in ATCGbio Life Technology Inc. is developed to be suitable for the follows:
1. Short (~15 bp) homologous recombination
2. Fragments of length from less than 100 bp to 6kb
3. Assembling multiple fragments. 
It is based on 5’->3’ exonuclease activity and is stable at -20 °C for more than 3 months. The cloning reaction requires only 20 minutes at 50 °C with 10 µl in volume. 
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