Molecular approaches for analyzing differential gene expression: differential cDNA library construction and screening.
Complementary deoxyribonucleic acid (cDNA) libraries can be used as a means to isolate and identify cell-specific messenger ribonucleic acid (mRNA) sequences. The basic elements of cDNA library construction and screening are reviewed in the context of analyzing differentially expressed mRNAs. A brief overview of the recombinant DNA systems applied to cDNA library construction and the principles of screening cDNA libraries by plaque hybridization are provided. Methods for comparing mRNA populations by differential screening and by competition hybridization are discussed, and methods for constructing subtracted cDNA libraries, enriched in differentially expressed sequences, are presented. Also reviewed are the analysis of differentially expressed cDNAs by Southern and Northern hybridization, RNase protection, polymerase chain reaction, and sequencing.
Calvet
J P
JP
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger; D004247:DNA