J Integr Plant Biol. ›› 1998, Vol. 40 ›› Issue (11): -.

• Research Articles •    

Studies on Microdissection and Microcloning of the Rye Chromosome 1R

JIANG Ci-Zhong, SONG Wen-Qin, LI Xiu-Lan and CHEN Rui-Yang   

Abstract: Two 1R chromosomes of Secale cereale L. were isolated from one metaphase cell by means of chromosome micro-isolation, and the chromosomal DNA was amplified adopting the cohesive adapters single primer polymerase chain reaction (CASP-PCR) technique. The CASP-PCR products were labeled as probes. The results of Southern blot hybridization confirmed that the CASP- PCR products derived from the chromosome IR were homologous with the genomic DNA of S. cereale. The clones of PCR products were obtained with high efficiency. Over 10 000 recombinant clones were obtained from one-tenth of the ligation mixture which was transferred into the competent E. coli DH5a. The size of the inserted fragments of clones ranged from 250 bp to 500 bp. This research has established the foundation for further selection of chromosome 1R markers.

Key words: Secale cereale, Chromosome 1R, Microdissection and microcloning, Cohesive adapters single primer polymerase chain reaction

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