Restriction Fragment Length Polymorphism: Image Analysis and Molecular Weight Calculation with a Scanner-Based Computer System
Image analysis and calculation programs are essential to enable precise DNA profiling and to reduce the subjective aspects of band measurements. We present a new system, called RFLP-MAC, Restriction Fragment Length Polymorphism — Molecular weight Analysis and Calculation. The system consists of a scanner unit and an Apple Macintosh computer supported by a combination of a shareware and self-developed software package. Molecular weights are calculated profiling the optical density distribution of each band. The migration distances of known marker fragments can be determined by the use of different mathematical models and gel distortions are corrected by a polynomial regression algorithm. All data are stored in a relational database system. Descriptive statistics can be performed as well as allele frequencies.
Serial intergel measurements are investigated in order to check the precision of the RFLP-MAC system; these data are subjected to a standard analysis of variance (maximal 3 SD =0.030 =1.723%). Allele size distribution of 380 unrelated individuals at the locus D10S28 (TBQ7, Hae Ill) are presented.
KeywordsMigration Agarose Electrophoresis
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