Enrichment of microsatellites from the citrus genome using biotinylated oligonucleotide sequences bound to streptavidin-coated magnetic particles.
A method is described for the rapid isolation of microsatellite sequences using a biotin-labeled oligonucleotide attached to streptavidin-coated magnetic particles. The oligonucleotide "hook" in solution hybridizes to complementary single-stranded lengths of genomic DNA onto which have been engineered specific PCR priming sites. The final product is an enriched library of microsatellites of defined sequences. The method is applicable to any genome and in principle is adaptable to the rapid isolation of both repetitive as well as genic sequences. It is illustrated by the isolation of trinucleotide repeat (TAA)n sequences from the citrus genome.
School of Biological Sciences, Flinders University of South Australia.
J M
JM
Kijas
J C
JC
Fowler
C A
CA
Garbett
M R
MR
Thomas
D001426:Bacterial Proteins; D001710:Biotin; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D017343:Genes, Plant; D008163:Luminescent Measurements; D008280:Magnetics; D008863:Microspheres; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D019809:Streptavidin
D001426:Bacterial Proteins; D009841:Oligonucleotides; D001710:Biotin; D004247:DNA; D019809:Streptavidin; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases