Colony hybridization to screen for microsatellites.
Simple sequence length polymorphisms (SSLPs) for use as genetic markers are derived from the microsatellite repeat sequences found abundantly throughout eukaryotic genomes. Microsatellite DNA is identified in libraries of cloned DNA by colony hybridization to oligonucleotide probes containing simple sequence repeat (SSR) DNA, a common procedure in molecular biology laboratories. Plasmid or phagemid colonies are transferred to a nylon filter, grown briefly, fixed, and hybridized with one or more 32P-labeled oligonucleotide probes. This protocol is adapted for the rapid processing of many filters at once and optimized for identification of long stretches of uninterrupted repetitive DNA sequences.
Hudson
T J
TJ
Whitehead Institute, Cambridge, Massachusetts, USA.
D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D015723:Gene Library; D005819:Genetic Markers; D005821:Genetic Techniques; D005826:Genetics, Medical; D006801:Humans; D018895:Microsatellite Repeats; D009693:Nucleic Acid Hybridization / Q000379:methods; D015345:Oligonucleotide Probes
D005819:Genetic Markers; D015345:Oligonucleotide Probes; D004247:DNA