High densities of unamplified single molecules on a surface can be extended asynchronously, thereby allowing substantial flexibility in the kinetics of sequencing chemistry. Single-molecule sequencing is an attractive approach due to its simplicity and the lack of cloning or amplification in sample preparation. Published results of various forms of single-molecule sequencing 11– 13 have shown improvements in throughput of about tenfold per year over the last few years, and no fundamental limits have yet been reached ( Supplementary Fig. 10), there has been rapid progress in the field. Since the first demonstration of single-molecule sequencing in 2003 (ref. Here we demonstrate that genome science is rapidly advancing to the point where individual instruments can achieve throughput that just a few years ago required the facilities of large genome centers. Reagent costs are substantial, estimated at $250,000–$500,000 per genome ( Supplementary Table 1).

Even using current next-generation technologies, however, sequencing of a human genome has required at least 35–40 machine runs with many instruments operating in parallel. Since the publication of the first rough draft consensus human genomes 1, 2, there have been several reports of individual human genome sequences 3– 9. There is broad interest in using human genome sequencing to better understand human genetic variation and genome-related diseases, such as cancer, and ultimately to guide discoveries and decisions about the health of individuals. This milestone should allow widespread application of genome sequencing to many aspects of genetics and human health, including personal genomics. We identified 752 regions of copy number variation by analyzing coverage depth alone and validated 27 of these using digital PCR. We determined ~2.8 million single nucleotide polymorphisms (SNPs) with a false-positive rate of less than 1% as validated by Sanger sequencing and 99.8% concordance with SNP genotyping arrays. Single-molecule sequencing enabled analysis of human genomic information without the need for cloning, amplification or ligation. Our results were obtained on one sequencing instrument by a single operator with four data collection runs. We aligned billions of 24- to 70-bp reads (32 bp average) to ~90% of the National Center for Biotechnology Information (NCBI) reference genome, with 28× average coverage. Here we report the use of single-molecule methods to sequence an individual human genome. Recent advances in high-throughput DNA sequencing technologies have enabled order-of-magnitude improvements in both cost and throughput.

In5d number sequences