The use of fluorescence detection and internal lane standards to size PCR products automatically

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 c...

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Bibliographic Details
Published inApplied and theoretical electrophoresis Vol. 3; no. 1; p. 1
Main Authors Mayrand, P E, Corcoran, K P, Ziegle, J S, Robertson, J M, Hoff, L B, Kronick, M N
Format Journal Article
LanguageEnglish
Published United States 1992
Subjects
Base Sequence
DNA - genetics
DNA - isolation & purification
Electrophoresis, Agar Gel
Female
Fluorescent Dyes
Genetic Carrier Screening
Humans
Male
Molecular Sequence Data
Molecular Weight
Muscular Dystrophies - genetics
Pedigree
Polymerase Chain Reaction - instrumentation
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - standards
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Summary: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.
ISSN:0954-6642