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.
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.
Trilink Web page： http://www.trilinkbiotech.com/cleantag/
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