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Design and synthesis of novel nucleotide analogs and protein conjugates for DNA sequencing

Wenjing Guo

Sequencing by Synthesis (SBS), a DNA sequencing methodology based on the DNA polymerase reaction, is a promising paradigm for deciphering large-scale genomes. This thesis describes the design and synthesis of a variety of nucleotide reversible terminators (NRTs) with different characteristics. One set of NRTs possesses a phosphate moiety attached to the 2’ position of the sugar to block further incorporation in polymerase reaction, with the potential for fluorescent tag attachment at the same site or on the base through a cleavable linker for detection. The other set of NRTs possesses an azido-methyl moiety that blocks the 3’-hydroxyl group for detection by surface-enhanced Raman scattering. Each NRT has been tested in proof-of-principle SBS experiments. In addition, a set of 5’-phosphate tagged nucleotides has been developed and tested for nanopore electronic detection. A new set of...

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