University of California

Christopher Vollmers

Charles Cole

Richard E Green

Theron Palmer

Roger Volden

Provided are methods of nucleic acid sequencing. The methods include producing a circularized DNA including a full-length cDNA and a known heterologous sequence, and performing rolling circle amplification using the circularized DNA as template to produce a concatemer including repeating segments including the full-length cDNA and the known heterologous sequence. The methods further include obtaining a raw sequencing read of the concatemer using a nanopore, identifying the repeating segments in the raw sequencing read, and producing a consensus sequence of the full-length cDNA based on the sequences of the repeating segments. Computer-readable media, computing devices, and systems that find use, e.g., in practicing the methods of the present disclosure are also provided.

Nucleic acid sequencing methods and computer-readable media for practicing same

Aspects of the present disclosure include methods of producing nucleic acid libraries. In certain aspects, the methods include producing tagged primer extension products, and contacting aliquots of the tagged primer extension products with transposomes to produce tagged extension product fragments. The methods may further include sequencing the tagged extension products and tagged extension product fragments to determine the sequences of nucleic acids of interest. Also provided are compositions and kits that find use, e.g., in practicing embodiments of the methods.

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Santa Cruz, CA, United States of America

Christopher Vollmers