PMC:3475479 / 4059-6169 JSONTXT

{"project":"2_test","denotations":[{"id":"23105922-16056220-44845627","span":{"begin":333,"end":334},"obj":"16056220"},{"id":"23105922-22192763-44845628","span":{"begin":1078,"end":1079},"obj":"22192763"},{"id":"23105922-22192914-44845629","span":{"begin":1081,"end":1082},"obj":"22192914"},{"id":"23105922-22247774-44845630","span":{"begin":1123,"end":1124},"obj":"22247774"},{"id":"23105922-21966408-44845631","span":{"begin":1126,"end":1127},"obj":"21966408"},{"id":"23105922-20398277-44845632","span":{"begin":1188,"end":1189},"obj":"20398277"},{"id":"23105922-15143316-44845633","span":{"begin":1359,"end":1360},"obj":"15143316"},{"id":"23105922-17055251-44845634","span":{"begin":1362,"end":1363},"obj":"17055251"},{"id":"23105922-17055251-44845635","span":{"begin":1962,"end":1963},"obj":"17055251"}],"text":"Current Next-generation Sequencing (NGS) Technology\n\nRoche/454 pyrosequencing technology\nThe first high-parallel sequencing system was developed with an emulsion PCR method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized on the individual well of a PicoTiterPlate (PTP) [1]. The DNA sequencing protocol, including sample preparation, is supplied for a user to follow the method, which is developed by 454 Life Science (now part of Roche Diagnostics, Mannheim, Germany). Even following the manufacturer's protocol, to obtain high-quality DNA sequence with maximal total DNA sequence length as possible, one should make a library with both adapters on the sheared DNA fragment and mix the optical ratio of library DNA versus bead for emulsion PCR. Now, the system is upgraded to through-put a total of an average of 700 Mbp with an average 600 bp/read in one PTP run. The sequencer is available for single reads and paired-end read sequencing for the application of eukaryotic and prokaryotic whole-genome sequencing [2, 3], metagenomics and microbial diversity [4, 5], and genetic variation detection for comparative genomics [6].\n\nIllumina/Solexa technology\nThe numerous cost-effective technologies were being developed for human genome resequencing that can be aligned to the reference sequence [7, 8]. The first successful technology to gain massive DNA sequencing available for resequencing was developed by Solexa (now part of Illumina, San Diego, CA, USA). The principle of the system is the method of base-by-base sequencing by synthesis, where the sheared template DNA is amplified on the flat surface slide (flow cell) and detects one base on each template per cycle with four base-specific, fluorescently labeled signals. Signals for all four fluorescent channels are collected and plotted at each position, enabling quality per scores to be derived using four-color information if desired [8]. Now, a maximum of 300 Gb for reading is available with 101 bp paired-end reading per fragment on a 1-flow cell run in the upgraded HiSeq system."}