A method of single chromosome isolation and amplification in Lycoris radiata was established.Root tip fixed by Carnoy fixative was digested with an enzyme mixture of cellulase and pectolyase,then used to prepare chromosome samples.The target chromosome was microdissection by using a hand made capillary needle under an inverted microscope.Each Eppendorf tube contained one chromosome.The dissected Lycoris radiata chromosomes were digested by proteinase K in 0.2 mL Eppendorf tubes respectively.After degenerating oligonucleotide primed PCR amplification(DOP PCR),DNA fragments were acquired.The size of DNA fragments in PCR products varied from 100 to 5 000 bp.This work would facilitate to construct the genomic DNA library of single chromosome and to screen the specific probes of single chromosome in Lycoris radiata.