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CleanTag® Library Prep

CleanTag Small RNA Library Prep Kit

PLOS publication
White Paper


CleanTag® Library Preparation for Next-Generation Sequencing

TriLink’s CleanTag technology solves one of the most pervasive problems in library preparation for next-generation sequencing (NGS), adapter dimer formation. Currently, many workflows involve a laborious gel purification step to rid the samples of the adapter dimer. TriLink applied its nucleic acid expertise to develop CleanTag modified adapters to block adapter dimer formation. This new technology:
  • Increases mappable sequencing reads even with ultra
    low RNA inputs
  • Tags efficiently in samples with low miRNA abundance
  • Suited for diagnostic samples with limited RNA
  • Eliminates the need for gel purification
  • Enables full automation
  • Compatible with Illumina and Ion Torrent* platforms

*Please contact our team for details when using with the Ion Torrent platform.

Now Available for Small RNA Library Preparation

Click to view: PLOS publication

CleanTag Technology

Adapter dimers form when the 5΄ and 3΄ adapters self-ligate without the target library insert. Consequently, amplification of this smaller adapter dimer side product often out-competes amplification of the tagged library. Our modified adapters block adapter-adapter ligation thus improving the adapter dimer to tagged library ratio. This leads to significantly higher quality sequencing data.

CleanTag Adapters Reduce Adapter Dimers While Maintaining Mapped Reads

Human total brain RNA was prepped using either TruSeq® Small RNA Sample Prep Kit or CleanTag Ligation
Kit for Small RNA Library Prep.


CleanTag Produces Superior Results with Samples Containing Low miRNA Abundance, such as Patient Blood and Plasma Samples

Human plasma samples contain a low abundance of small RNAs and have traditionally been difficult to sequence. In collaboration with Hudson Alpha Institute for Biotechnology, we show that the CleanTag Ligation Kit for Small RNA Library Prep efficiently tags small RNA and produces significantly more valuable reads when compared to a previously published protocol optimized for adapter dimer reduction.

Adaptor-Dimer Reduction in microRNA-seq library Sequence Read Disposition. 1 uL RNA isolated from 1 mL of human plasma (~20ng) was used to generate microRNA-seq libraries using the protocol published by Vigneault, et al. (2012) and the CleanTag Small RNA Protocol. The two protocols were performed separately and combined onto one gel separation step, and microRNA bands (~140bp) were excised, processed and sequenced with 50bp single end protocol on the Illumina MiSeq Platform (total ~15 Million Reads). CleanTag modied adapters were diluted 1:4 and 15 PCR cycles were performed.

CleanTag Eliminates Need for Gel Purification

1000 ng Human total brain RNA input. AMPure® XP purified. Analyzed by BioAnalyzer®.


CleanTag Renders Small RNA-Seq Fully Automatable

The size of the adapter dimer and tagged library are similar in length, thus requiring a gel size selection step to enrich the libraries for the desired product. Because of this, a fully automated workflow was not possible. Now, with TriLink's CleanTag technology, there is no for need gel purification, thus allowing full automation of small RNA-seq for the first time.

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