Automated polymerase chain reaction product sample preparation for capillary electrophoresis analysis

The analysis of crude polymerase chain reaction (PCR) products by capillary electrophoresis (CE) is often compromised due to the presence of a high concentration of salt. Salt interferes with the electrokinetic injection and induces localized heating within the column; hence, PCR products must be de...

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Bibliographic Details
Published in:Journal of chromatography. B, Biomedical applications Biomedical applications, 1996-08, Vol.683 (1), p.109-114
Main Authors: Belgrader, Phillip, Devaney, Joseph M., Del Rio, Susie A., Turner, Kelly A., Weaver, Kristal R., Marino, Michael A.
Format: Article
Language:English
Subjects:
Automation
DNA - analysis
Electrophoresis, Capillary - methods
Enzymes
Polymerase chain reaction
Polymerase Chain Reaction - methods
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Summary:The analysis of crude polymerase chain reaction (PCR) products by capillary electrophoresis (CE) is often compromised due to the presence of a high concentration of salt. Salt interferes with the electrokinetic injection and induces localized heating within the column; hence, PCR products must be desalted or cleaned-up prior to CE analysis. A variety of commercial clean-up systems are available that have been traditionally used to prepare PCR products for cloning, sequencing and digestion with restriction enzymes. These systems were tested for their effectiveness in preparing PCR products for CE analysis and were evaluated based on CE resolution, salt removal, DNA recovery, processing time and cost. One particularly effective clean-up system, membrane dialysis, was automated using a robotic workstation.
ISSN:0378-4347
1572-6495
DOI:10.1016/0378-4347(96)00165-X