Diagenode

Directional high-throughput sequencing of RNAs without gene-specific primers.


Mäki A, Tiirola M

Ribosomal RNA analysis is a useful tool for characterization of microbial communities. However, the lack of broad-range primers has hampered the simultaneous analysis of eukaryotic and prokaryotic members by amplicon sequencing. We present a complete workflow for directional, primer-independent sequencing of size-selected small subunit ribosomal RNA fragments. The library preparation protocol includes gel extraction of the target RNA, ligation of an RNA oligo to the 5'-end of the target, and cDNA synthesis with a tailed random-hexamer primer and further barcoding. The sequencing results of a phytoplankton mock community showed a highly similar profile to the biomass indicators. This method has universal potential for microbiome studies, and is compatible for the 5'-end sequencing of other RNA types with minimum library preparation costs.

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Published
October, 2018

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    C05010041
    CATS RNA-seq Kit v2 x24

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