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Automated polymerase chain reaction product sample preparation for capillary electrophoresis analysis
P Belgrader1, J M Devaney, S A Del Rio
1Advanced Technologies Development Branch, Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD 20850, USA.
Abstract:
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.
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