石蒜单染色体的显微分离及体外扩增

建立了石蒜单染色体的分离及体外扩增的方法。石蒜根尖经卡诺固定液固定后，再用果胶酶和纤维素酶酶解处理，制得标本，在倒置显微镜下，用自制的毛细管针挑取目的的染色体。将分离的石蒜染色体放入0．2mLEppedorf管中，经蛋白酶K处理后，进行DOP－PCR扩增，获得DNA片段。琼脂糖凝胶电泳显示扩增产物的长度大约为100－5000bp。此法为构建石蒜单染色体基因组库和筛选其特异性探针奠定了基础。

Microdissection and PCR Amplification of Single Chromosome of Lycoris radiata

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.