title: Simple and effective method for generating single-stranded DNA targets and probes.
abstract: A simple and efficient PCR method was developed for generating dye- or radiolabeled single-stranded DNA targets or probes used for hybridization studies. The method involved the use of a pair of long primers with high annealing temperatures and a short, labeled primer with a low annealing temperature in a PCR consisting of two cycles at different temperatures. We used this method to generate dye Cy 5-labeled and [32P]-radiolabeled single-stranded DNA targets and probes. These labeled probes were used successfully for the microarray identification of point mutations in Mycobacterium tuberculosis genes and for the Northern blot detection of expression changes of the GATA-2 gene in Pneumocystis carinii-infected rat lungs.
authors.affiliation: U.S. Food and Drug Administration, Rockville, MD, USA. xxt4@cdrh.fda.gov
authors.forename: Xing
authors.initials: X
authors.lastname: Tang
authors.forename: Sheldon L
authors.initials: SL
authors.lastname: Morris
authors.forename: John J
authors.initials: JJ
authors.lastname: Langone
authors.forename: Larry E
authors.initials: LE
authors.lastname: Bockstahler
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D017931:DNA Primers; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000235:genetics*; D008168:Lung / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D051381:Rats
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004277:DNA, Single-Stranded