Next-generation sequencing (NGS) platforms achieve massively parallel sequencing. The Illumina platform uses a sequencing-by-synthesis approach that employs which terminator chemistry to achieve single-base resolution with reversible stopping?

• A Dideoxy chain terminators (ddNTPs) that permanently block chain extension
• B Reversible dye-labeled 3'-blocked nucleotide terminators ✓
• C Pyrophosphate detection from natural nucleotide incorporation (pyrosequencing)
• D Nanopore electrical current changes as single-stranded DNA threads through a pore

Explanation

Illumina sequencing (sequencing by synthesis) uses fluorescently labelled 3'-blocked reversible terminators: each cycle, one nucleotide is incorporated, synthesis stops (due to 3' block), the fluorescent signal is imaged to identify the base, and then the 3' block is chemically removed to allow the next cycle. This achieves single-base resolution across millions of clusters simultaneously. Dideoxy terminators are used in Sanger sequencing (irreversible); pyrophosphate detection is used in 454/Roche pyrosequencing; nanopore electrical current is Oxford Nanopore Technology for long-read sequencing.

Reference: Harper's Illustrated Biochemistry, 32nd ed.

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