Optimization of polymer replacement protocol for novel automated single-lane DNA sequencing instrument based on single photon detection

نویسندگان

  • Olga Bilenko
  • Dmitri N. Gavrilov
  • Boris Gorbovitski
  • Mikhail Gouzman
  • Georgiy Gudkov
  • Vyacheslav Khozikov
  • Olga Kosobokova
  • Nadia Lifshitz
  • Serge Luryi
  • Andrew Stepoukhovitch
  • Marina Tcherevishnick
  • Andrei Tsuprik
  • Georgiy Tyshko
چکیده

An automated ultra-sensitive single-lane DNA sequencer based on single photon counting fluorescent detection technique has been developed and tested. The instrument is intended for individual researchers who carry out a limited number of sequencing experiments (about 10 bp/year, the size typical for small-scale research projects). Due to the use of single photon detection, sensitivity of the instrument is about 100 fold higher than in commercial ABI instruments. The sequencer has a flexible modular fiber-optic design and can be easily configured to operate with different laser sources, as well as various fiberized photodetectors. A novel automation concept providing the ability of the remote (Internet) access and control of the system has been implemented and tested for DNA sequencing and fragment analysis applications. Optimization of sample running protocol enabled up to 12 hours of continuous operation without polymer replacement for standard ABI sequencing chemistry and reagents.

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تاریخ انتشار 2003