Next-Generation Sequencing (NGS)

Next-generation sequencing (NGS) has revolutionized the sequencing of nucleic acids, as it can usually yield millions or tens of millions of reads per experiment. Moreover, the cost of NGS is rapidly falling, which has resulted in an explosion of data sequencing. Sequencing of small RNAs, such as microRNAs (miRNAs), tRNA-derived fragments (tRFs), and others, has also gained popularity. miRNAs are small ~22 nucleotide fragments that regulate families of mRNAs by binding to their 3’ untranslated regions. miRNA regulation has been implicated in differentiation, cell fate, cancer, and numerous other disease states, making miRNAs popular biomarkers. However, sequencing of miRNAs and other small RNAs is no easy feat. In common approaches, such as Illumina® or Ion Torrent™ small RNA sequencing, 5’ and 3’ adapters are ligated to the target RNA to create a library. Unfortunately, the 5’ and 3’ adapter can also ligate together in the absence of an intervening RNA sequence, forming adapter dimers. Because the size of the library is only slightly larger than the adapter dimer, most protocols require a gel purification step to reduce the adapter dimer. At very low RNA inputs, the adapter dimer dominates the sequencing reads and prohibits sequencing. To overcome this issue, TriLink developed the CleanTag® Small RNA Library Prep Kit. The CleanTag kit uses chemically modified adapters that inhibit adapter dimers. This allows for efficient sequencing of low input samples, such as biofluids, without the need for a gel purification step. Recently, in collaboration with leading researchers in the field, we were also able to efficiently sequence small RNAs from single cells. In addition, TriLink NGS products include Illumina® and Ion Torrent™ index primers for your NGS experiments.

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