6533b821fe1ef96bd127c321
RESEARCH PRODUCT
Nonradioactive Detection of Differentially Expressed Genes Using Complex RNA or DNA Hybridization Probes
Ralf RossAngelika B. Reske-kunzT. LaenginB. RuegerXiao-lan Rosssubject
MaleCandidate geneDNA ComplementaryMolecular Probe TechniquesBiologySensitivity and SpecificityGeneral Biochemistry Genetics and Molecular BiologyMiceDigGene expressionAnimalsHumansGeneGenomic LibraryMice Inbred BALB CMessenger RNADNA–DNA hybridizationNucleic Acid HybridizationRNARNA ProbesMolecular biologyGene Expression RegulationGenesLangerhans CellsLuminescent MeasurementsFemaleMolecular probeDigoxigeninBiotechnologydescription
The analysis of differential gene expression has become increasingly important in recent years. Typically, differentially expressed genes are identified in a primary screening procedure, yielding candidate genes whose differential expression has to be verified. We provide a highly sensitive, efficient and nonradioactive differential screening procedure to analyze numerous candidate genes in a single step. This comprises labeling of poly(A)+ RNA of the cell types analyzed with DIG Chem-Link and differential hybridization to the candidate genes fixed on dot blots. DIG Chem-Link allows, to our knowledge, for the first time efficient and direct nonradioactive labeling of RNA in vitro. Advantages of this method include extremely short exposure times and the feasibility to re-use the probes after prolonged storage. Using this procedure, we isolated several genes that are differentially expressed in maturing Langerhans cells.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1999-01-23 | BioTechniques |