Issue Archive

Cloning A microdissected chromosome of soybean and in situ analysis of the chromosome PCR products

Zhou Yihua, Hu Zanmin, Wang Lanlan, Chen Zhenghua

Institute of Genetics,Chinese Academy of Sciences.Beijing 100101

Abstract

A single chromosome was microdissected from the metaphase chromosomes of soybean by using a glass needle. After two rounds of Sau3A linker-adaptor-mediated PCR, the second-round PCR products from the microdissected chromosome were cloned into plasmid vector to generate a chromosome-specific microclone library, which included approximately 200 000 recombinant clones. 178 randomly selected clones were estimated to be ranging in size from 200-1 800bp, with an average size of 830bp, of which approximately 44% were high/medium copy clones, while 56% were low/unique copy clones. In situ hybridization of the chromosome PCR products to the mitotic metaphase spreads showed that the PCR products were hybridized to the genomic soybean DNA, but the labeling was not restricted to the microsissected chromosome.

Key words: Microdissection;Microcloning;A single chromosome;Soybean

Received: 　　Accepted:

Corresponding author: 　　E-mail:

Citing This Article：

Zhou Yihua, Hu Zanmin, Wang Lanlan, Chen Zhenghua. Cloning A microdissected chromosome of soybean and in situ analysis of the chromosome PCR products. Acta Physiol Sin 2000; 52 (3): (in Chinese with English abstract).