Chromosome microdissection by laser microbeam, chromosomal fragment isolation and amplification in vitro in barley (Hordeum vulgare L.)

作     者：Huai Wang Yihua Zhou Benyuan Dang Zanmin Hu Lanlan Wang Liangcai Li Zhenghua Chen 

作者机构：Acad Sinica Inst Genet Beijing 100101 Peoples R China 

出 版 物：《Chinese Science Bulletin》 (Chin. Sci. Bull.)

年 卷 期：1998年第43卷第10期

页      面：851-855页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 090102[农学-作物遗传育种] 

主　　题：Hordeum vulgare L. chromosome microdissection laser microbeam LA_PCR. 

摘      要：A method for microdissection, isolation and amplification of plant chromosomal fragments using laser microbeam and a glass microneedle was established. Firstly, 7H chromosome of barley (Hordeum vulgare L.) was dissected by Nd: YAG laserbeam with suitable parameters and the fragment comprising a satellite was isolated with a glass microneedle which was fixed on a micromanipulator. Then, the chromosomal fragment DNA was amplified by LA_PCR (linker adaptor PCR) for two rounds. The size of the DNA fragments of PCR products varied from 500-3 000 bp and the PCR products originated from the genome of barley were verified by Southern hybridization. Compared with previous reports, there are some advantages in this research. The performance is easier, the dissection is more precise and the cost is low. It also permits efficient amplification with only one single chromosome fragment. Laser microbeam_glass microneedle method may be useful in the microdissection of special chromosome regions, especially in plants with middle or small chromosomes.