ABSTRACT
Recently, we developed a family of high-performance automated capillary DNA sequencing instruments based on a single-photon detection of fluorescently labeled DNA fragments. Our machines employ digital and broadband techniques, essential for achieving superior instrument sensitivity and dynamic range. In the present paper, we discuss limitations of the instrument's performance caused by the nonlinearity of single-photon detectors as well as methods for nonlinearity compensation which increase the detection dynamic range and base-calling accuracy.
Subject(s)
Electrophoresis, Capillary/instrumentation , Nucleotides/analysis , Sequence Analysis, DNA/instrumentation , Electronics , Electrophoresis, Capillary/standards , Equipment Design , Fluorescent Dyes/analysis , Photons , Reference Values , Reproducibility of Results , Sequence Analysis, DNA/standardsABSTRACT
We have developed a family of high-performance capillary DNA sequencing instruments based on a novel multicolor fluorescent detection technology. This technology is based on two technical innovations: the multilaser excitation of fluorescence of labeled DNA fragments and the "color-blind" single-photon detection of modulated fluorescence. Our machines employ modern digital and broadband techniques that are essential for achieving superior instrument performance. We discuss the design and testing results for several versions of the automated single lane DNA sequencers, as well as our approach to scaling up to multilane instruments.
