title: General method for HPLC purification and sequencing of selected dsDNA gene fragments from complex PCRs generated during gene expression profiling.
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.
authors.affiliation: Celera Genomics, Foster City, CA, USA.
authors.forename: L Y
authors.initials: LY
authors.lastname: Wong
authors.forename: V
authors.initials: V
authors.lastname: Belonogoff
authors.forename: V L
authors.initials: VL
authors.lastname: Boyd
authors.forename: N M
authors.initials: NM
authors.lastname: Hunkapiller
authors.forename: P M
authors.initials: PM
authors.lastname: Casey
authors.forename: S N
authors.initials: SN
authors.lastname: Liew
authors.forename: K D
authors.initials: KD
authors.lastname: Lazaruk
authors.forename: S
authors.initials: S
authors.lastname: Baumhueter
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D005069:Evaluation Studies as Topic; D020869:Gene Expression Profiling / Q000379:methods*; D008099:Liver / Q000737:chemistry; D016133:Polymerase Chain Reaction; D051381:Rats; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004247:DNA