Differential gene expression analysis by micro-preparative capillary gel electrophoresis.
Differential display analysis by cDNA fractionation, collection of differentially expressed fractions of interests and their downstream characterization is demonstrated. cDNA pools from two strains of Cochliobolus heterostrophus fungus were generated by specific restriction digestion and selective ligation. Micropreparative separation and isolation of differentially expressed transcript representatives were accomplished by high-performance capillary gel electrophoresis. The collected individual DNA molecules were polymerase chain reaction amplified and sequenced to create expressed sequence tags for the genes of interests. High resolving power and sensitivity of capillary gel electrophoresis enabled fast and automated processing of minute amounts of cDNA samples with high precision.
Guttman
Andras
A
Torrey Mesa Research Institute, 3115 Merryfield Row, San Diego, CA 92121, USA. aguttman@diversa.com
Shi
Liang
L
Khandurina
Julia
J
Wang
Xun
X
D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary; D019075:Electrophoresis, Capillary / Q000379:methods*; D020869:Gene Expression Profiling; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018076:DNA, Complementary