tRNA can be sequenced with nanopore sequencers, so long as they can be unfolded and electrically attracted to the nanopore. So a mechanism to capture tRNA molecules, unfold them, and initiate threading them into a nanopore is needed.
The invention includes attaching DNA or RNA "handles" to a tRNA molecule. These handles allow manipulation of the tRNA molecule, including unfolding its structure and acting as targets for attaching other molecules to the tRNA.
One example is a double stranded oligonucleotide adapter that can be ligated to a tRNA from a biological sample. Such an adepter can be a Y shaped double stranded DNA-RNA adapter with a 3' RNA overhang complementary to the CCA tail present in tRNA. The adapter can also include a cholesterol tag within its 3' end.
Examples of sequences used in oligonucleotide adaptors include:
X indicates an abasic 1'2' dideoxyribose; Z indicates a triethylene glycol cholesterol
Molecular adaptor that facilitates tRNA seqeuncing by nanopore
Rapid identification of tRNA species in a biological sample
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tRNA sequencing, sequencing adapter, tRNA, nanopore sequencing, RNA, RNAseq, long read sequencing