Compensation for mobility inequalities between lanes computed from band signals in on-line fluorescence DNA sequencing.
Among on-line fluorescence DNA sequencing systems, the four-lane method exhibits the potential for reporting an erroneous sequence due to nonuniform mobility of the DNA fragments migrating among the four lanes. This error is manifest in phenomenon commonly called smiling. This paper presents a computational algorithm which compensates for the mobility inequalities between lanes using signal data obtained from the shorter DNA fragments forming the faster migrating bands. The program mainly consists of two routines: (i) calculation of calibration coefficients (mobility ratios between lanes), and (ii) examination of the coefficients by applying them to a later domain of the same signals. Both routines are connected with several feed-back branches for recalculation. Homology analysis of final sequences has shown that the accuracy rate is maximized with this algorithm and any ambiguous result can be assigned to the residual error inherent in the band identification method used.
Fujii
H
H
Biotechnology Instruments Department, Shimadzu Corporation, Kyoto, Japan.
Kashiwagi
K
K
D000465:Algorithms; D001483:Base Sequence; D002138:Calibration; D004247:DNA / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted; D012984:Software; D013050:Spectrometry, Fluorescence
D004277:DNA, Single-Stranded; D004247:DNA