Abstract
cDNA microarray technology is becoming the technique of choice for studying gene expression and gene expression patterns. Although experimental protocols are available, only limited methodological information on microarray manufacture, hybridization, and signal interpretation has been published. The aim of this paper is to provide more insight into the practical aspects of microarray construction and hybridization. The influence of the size, composition, and concentration of the spotted DNA fragments on the final hybridization signal and the effect of hybridization volume, sample concentration, and sample depletion have been tested and are discussed.
MeSH terms
-
Amines / chemistry
-
Base Composition
-
Caco-2 Cells
-
Cloning, Molecular
-
DNA Primers / genetics
-
DNA Primers / metabolism
-
DNA, Complementary / genetics*
-
DNA, Complementary / metabolism
-
Epithelial Cells / metabolism
-
Genetic Vectors / metabolism
-
Glass / chemistry*
-
Humans
-
Intestinal Mucosa / cytology
-
Luciferases / genetics
-
Luciferases / metabolism
-
Oligonucleotide Array Sequence Analysis / methods*
-
Petunia / metabolism
-
Poly A / metabolism
-
Polymerase Chain Reaction / methods
-
RNA, Messenger / metabolism
-
Sensitivity and Specificity
-
Silanes / chemistry*
-
Substrate Specificity
Substances
-
Amines
-
DNA Primers
-
DNA, Complementary
-
RNA, Messenger
-
Silanes
-
Poly A
-
Luciferases