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Title: Preparation of single rice chromosome for construction of a DNA library using a laser microbeam trap
Author: Liu, XH ; Wang, HW ; Li, YM ; Tang, YS ; Liu, YL ; Hu, X ; Jia, PX ; Ying, K ; Feng, Q ; Guan, JP ; Jin, CQ ; Zhang, L ; Lou, LR ; Zhou, ZA ; Han, B
Source: JOURNAL OF BIOTECHNOLOGY
Issued Date: 2004
Volume: 109, Issue:3, Pages:217-226
Keyword: optical scissors ; optical trap ; single rice chromosome ; adaptor PCR ; GENETICS ; CELLS ; TOOL
Subject: Biotechnology & Applied Microbiology
Corresponding Author: Han, B (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Natl Ctr Gene Res, Shanghai 200233, Peoples R China,bhan@ncgr.ac.cn
English Abstract: We report the development of a laser micromanipulation system and its application in the isolation of individual rice chromosomes directly from a metaphase cell. Microdissection and flow sorting are two major methods for the isolation of single chromosome. These methods are dependent on the techniques of chromosome spread and chromosome suspension, respectively. In the development of this system, we avoided using chromosome spread and cell suspension was used instead. The cell wall of metaphase rice cell was cut by optical scissors. The released single chromosome was captured by an optical trap and transported to an area without cell debris. The isolated single chromosome was then collected and specific library was constructed by linker adaptor PCR. The average insert size of the library was about 300bp. Two hundred inserts of chromosome 4 library were sequenced, and 96.5% were aligned to the corresponding sequences of rice chromosome 4. These results suggest the possible application of this method for the preparation of other subcellular structures and for the cloning of single macromolecule through a laser microbeam trap. (C) 2004 Elsevier B.V. All rights reserved.
Indexed Type: sci
Language: 英语
Content Type: 期刊论文
URI: http://ir.sibs.ac.cn/handle/331001/1965
Appears in Collections:神经所(总)_期刊论文

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Liu, XH; Wang, HW; Li, YM; Tang, YS; Liu, YL; Hu, X; Jia, PX; Ying, K; Feng, Q; Guan, JP; Jin, CQ; Zhang, L; Lou, LR; Zhou, ZA; Han, B.Preparation of single rice chromosome for construction of a DNA library using a laser microbeam trap,JOURNAL OF BIOTECHNOLOGY,2004,109(3):217-226
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