General method for HPLC purification and sequencing of selected dsDNA gene fragments from complex PCRs generated during gene expression profiling.
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.
Wong
L Y
LY
Celera Genomics, Foster City, CA, USA.
Belonogoff
V
V
Boyd
V L
VL
Hunkapiller
N M
NM
Casey
P M
PM
Liew
S N
SN
Lazaruk
K D
KD
Baumhueter
S
S
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*
D004247:DNA