LI-COR sequencers can routinely provide 800-1100 nucleotide, one-pass reads with accuracy better than 99% on single and double-stranded DNA templates. These instruments produce long nucleotide reads through the use of a focusing microscope mounted on a scanning laser to detect near-infrared fluorescence labeled products from DNA sequencing reactions. The number of nucleotides determined from a set of four lanes is a function of the quality of DNA template and the length of the gel used to resolve the reaction products. The high accuracy levels are, in part, attributable to the use of the four-lane sequencing strategy. Since raw data are de-convoluted (fluorescent labeled reaction products for each of the dideoxy-nucleotide chain terminations are electrophoresed in separate lanes) spectral overlap leading to incorrect or ambiguous base calls does not occur. The four lane presentation format also permits the rapid review and editing of sequence reads; however, this format allows the analysis of fewer reactions on a single gel.
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