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NGS Click ligation

enables efficient library preparation for RNA sequencing

Despite the variety and quality of existing sequencing methods, there is always an effort to improve them in every aspect (cost, speed, accuracy).
In the past year, baseclick has developed a click chemistry-based method for the non-enzymatic ligation of oligonucleotides, which is efficient, site-specific and is compatible with PCR amplification (see figure).
Moreover, the reaction between two non-templated single stranded oligonucleotides is more efficient than the comparable enzymatic ligation reaction, thus eliminating the need for second strand synthesis in e. g. RNA sequencing.

The preparation of alkyne- or azide-containing oligos has been studied extensively. Of special interest for this invention are “backbone mimics”, i. e. non-natural alternatives for the phosphodiester bond, which can be generated by copper-catalyzed azide alkyne cycloadditions (CuAAC). Some of the resulting triazole-containing oligonucleotides can be converted into natural phosphodiester backbones by polymerase enzymes without mutation of the sequence and thus are fully biocompatible.

Outline of the ClickAdapt workflow for NGS library preparation:



 
Especially useful applications of this artificial DNA backbone technology could be in sample preparation for next-generation sequencing.

Baseclick now sets out to open this emerging and highly potent technology to the R&D community by soon launching an NGS library preparation kit. Moreover, we have started strong partnerships to explore application opportunities for the different NGS technologies.  First start was already done by patent application of this outstanding methodology. Click here for more information on our patents.

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