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1. General method for HPLC purification and sequencing of selected dsDNA gene fragments from complex PCRs generated during gene expression profiling.

Author

Wong LY, Belonogoff V, Boyd VL, Hunkapiller NM, Casey PM, Liew SN, Lazaruk KD, and Baumhueter S

Subjects
Animals, Brain Chemistry, DNA analysis, Electrophoresis, Capillary, Evaluation Studies as Topic, Liver chemistry, Rats, Reproducibility of Results, Chromatography, High Pressure Liquid methods, DNA genetics, DNA isolation & purification, Gene Expression Profiling methods, Polymerase Chain Reaction, Sequence Analysis, DNA methods
Abstract

Gene expression profiling using an AFLP-based technique generates a large number of gene fragments that require identification by sequencing. The DNA fragments vary in length from about 50-500 bp. Ion-pair reversed-phase HPLC can be used to purify selected double-stranded DNA fragments that represent differentially expressed genes. The gene fragments are sequenced directly after vacuum drying of the collected HPLC fractions.

Published
2000
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