nanda runda

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+[[!meta title="Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs."]]
+[[!tag single_cell method transcriptomei ssbp]]
+
+Nat Commun. 2018 Feb 12;9(1):619. doi:10.1038/s41467-018-02866-0
+
+Hayashi T, Ozaki H, Sasagawa Y, Umeda M, Danno H, Nikaido I.
+
+Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs.
+
+[[!pmid 29434199 desc="First-strand cDNAs are synthesised with a RNAse H minus reverse-transcriptase.  DNAse I introduces nicks that prime synthesis of new cDNA molecules.  T4gp32 promotes the strand-displacement activity of the reverse-transcriptase.  Second-strand cDNAs are synthethised with Klenow fragment (3′ → 5′ exo-) primed with NSRs.  In single cells, genomic DNA needs to be removed because of the DNAseI digestion during RT and the low-complexity priming of the second-strand synthesis with NSRs.  The resulting DNA molecules are tagmented and sequenced with standard methods.  Thus, the method is non-stranded.  Despite the use of NSRs, the rRNA rate stays between 20 and 30 %."]]