title: The use of fluorescence detection and internal lane standards to size PCR products automatically.
abstract: We have developed chemical procedures, optical and electrophoretic instrumentation and computer software automate the analysis of polymerase chain reaction (PCR) products. DNA molecules labeled with up to four different fluorescent dyes are analyzed within a single electrophoresis gel lane. A size calibration curve is created for each electrophoresis lane from the electrophoretogram of uniquely labeled DNA fragments belonging to an internal lane standard that co-electrophoreses with the PCR products. The unknown molecular lengths of PCR products are automatically calculated from the calibration curve. Data from control experiments with DNA segments of known molecular length demonstrate the accuracy and precision of such sizing. This system has been applied to the analysis of PCR products for research in the areas of human identification, genetic mapping and genetic disease.
authors.affiliation: Applied Biosystems, Inc., Foster City, California.
authors.forename: P E
authors.initials: PE
authors.lastname: Mayrand
authors.forename: K P
authors.initials: KP
authors.lastname: Corcoran
authors.forename: J S
authors.initials: JS
authors.lastname: Ziegle
authors.forename: J M
authors.initials: JM
authors.lastname: Robertson
authors.forename: L B
authors.initials: LB
authors.lastname: Hoff
authors.forename: M N
authors.initials: MN
authors.lastname: Kronick
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005260:Female; D005456:Fluorescent Dyes; D006580:Genetic Carrier Screening; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009136:Muscular Dystrophies / Q000235:genetics; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods* / Q000592:standards
chemical_list: D005456:Fluorescent Dyes; D004247:DNA