Table 1.

Comparison of Commercially Available Next-Generation Sequencing Technologies Detailing Similarities and Differences

Technology	Library Construction	Sequencing Mechanism	Detection Mechanism	Maximum Read Length (bp)	Error Mode
First generation
ABI Sanger	Bacterial cloning	Dideoxy chain termination	Fluorescence	900	End-of-read errors
Massively parallel sequencing, part 1
Roche 454	Emulsion PCR on microbead surface	Polymerase-mediated incorporation of unlabelled nucleotides	Photon detection (light)	700	Errors in homopolymer runs
Illumina HiSeq	Amplification on glass surface	Polymerase-mediated incorporation of fluorescent nucleotides	Fluorescence	250	End-of-read errors
Life Technologies SOLiD	Emulsion PCR on microbead surface	Ligase-mediated addition of two base-encoded fluorescent oligonucleotides	Fluorescence	75	End-of-read errors
Massively parallel sequencing, part 2
Helicos	N/A (single molecule detection)	Polymerase-mediated incorporation of fluorescent nucleotides	Fluorescence	32	End-of-read errors
Life Technologies Ion Torrent	Emulsion PCR on microbead surface	Polymerase-mediated incorporation of unlabelled nucleotides	Ion sensing by semiconductors	200	Errors in homopolymer runs
Pacific Biosciences	N/A (single molecule detection)	Polymerase-mediated incorporation of fluorescent nucleotides	Fluorescence in real time	> 1,000	Random errors
Oxford Nanopore	N/A (single molecule detection)	Depolymerization and cleavage of individual nucleotides	Ion sensing by nanopore in electrically resistant membrane bilayer	Approximately 50	Errors generated by slipping or skipping of DNA

NOTE. Massively parallel sequencing has evolved in two stages: part 1 includes technologies that were developed before 2008; part 2 indicates technologies that have one or more characteristics that define an emerging technology (ie, single-molecule sequencing or direct detection of nucleotide signal).

Abbreviations: N/A, not applicable; PCR, polymerase chain reaction.