pattern via 65-μm-wide turns. Sequencing chemistry Thermal cycling In the TC reaction chamber, dye-terminator sequencing reagent, template DNA, and primers are loaded into the TC chamber and thermal-cycled for 35 cycles ( at 95 °C for 12 seconds and at 60 °C for 55 seconds). Purification The charged reaction mixture (containing extension fragments, template DNA, and excess sequencing reagent) is conducted through a capture/purification chamber at 30 °C via a 33-Volts/cm electric field applied between capture outlet and inlet ports. The capture gel through which the sample is driven, consists of 40 μM of oligonucleotide (complementary to the primers) covalently bound to a polyacrylamide matrix. Extension fragments are immobilized by the gel matrix, and excess primer, template, free nucleotides, and salts are eluted through the capture waste port. The capture gel is heated to 67-75 °C to release extension fragments. Capillary electrophoresis Extension fragments are injected into the CE chamber where they are electrophoresed through a 125-167-V/cm field. Platforms The Apollo 100 platform (Microchip Biotechnologies Inc., Dublin, CA) integrates the first two Sanger sequencing steps (thermal cycling and purification) in a fully automated system. The manufacturer claims that samples are ready for capillary electrophoresis within three hours of the sample and reagents being loaded into the system. The Apollo 100 platform requires sub-microliter volumes of reagents. Comparisons to other sequencing techniques The ultimate goal of high-throughput sequencing is to develop systems that are low-cost, and extremely efficient at obtaining extended (longer) read lengths. Longer read lengths of each single electrophoretic separation, substantially reduces the cost associated with de novo DNA sequencing and the number of templates needed to sequence DNA contigs at a given redundancy. Microfluidics may allow for faster, cheaper and easier sequence assembly. See also Second-generation sequencing Third-generation sequencing References Further reading External links MBI Says New Tool That Automates Sanger Sample Prep Cuts Reagent and Labor Costs DNA sequencing methods Molecular biology techniques 1977 in biotechnology de:Sanger-Methode