J Integr Plant Biol. ›› 1998, Vol. 40 ›› Issue (2): -.

• Research Articles •    

Microdissection and PCR Amplification of Single Soybean Chromosome

ZHOU Yi-Hua,DANG Ben-Yuan, HU Zan-Min,CUI Li-Hua, LI Liang-Cai and CHEN Zheng-Hua   

Abstract: Glass needles were successfully used to dissect the soybean (Glycine max L. ) single chromosome under the micromanipulator in this research. Two dissected soybean chromosomes were digested by Sau3A in two 0.5 mL Eppendorf tubes respectively. The two ends of chromosomal fragments were ligated with Sau3A linker adoptor. After two rounds of PCR amplification, smear DNA fragments ranged from 0.3 to 3 kb were acquired. Southern hybridization result showed the PCR products from the two single soybean chromosomes were homogeneous with the soybean genomic DNA, indicating that DNAs from the two single chromosomes have been successfully amplified. At the same time, the amplified products from the two of the distinguished single chromosome appeared somewhat different. The authors dissected the small chromosomes only by a traditional inverted microscope. Therefore, this research provides a plausible chance for amplification and microcloning of single small chromosomes.

Key words: Soybean, Microdissection, Single chromosome

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