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Cotton Science    2017, Vol. 29 Issue (3) : 213-221     DOI: 10.11963/1002-7807.lylprh.20170427
RESEARCH REPORTS |
Construction and Primary Application of Oligos Fluorescence in situ Hybridization Technology in Cotton
Yuling Liu1,2(),Zhen Liu1,Zhaoguo Li1,Yuhong Wang2,Zhongli Zhou2,Xiaoyan Cai2,Xingxing Wang2,Xiaoyan Wang1,Shulin Zhang1,Haiyan Zhao1,Zhenmei Zhang2,Kunbo Wang2,Fang Liu2,*(),Renhai Peng1,2,*()
1. Anyang Institute of Technology, Anyang, Henan 455000, China
2. Institute of Cotton Research of Chinese Academy of Agricultural Sciences / State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China
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Abstract  

[Objective] Fluorescence in situ hybridization (FISH) can achieve the accurate chromosomal localization of DNA sequences, which is one of the important techniques for genome in-depth study. The acquisition of chromosome-specific probes is the key for FISH application. The aim of this study is to explore a system of cotton oligonucleotides (oligos) fluorescence in situ hybridization (Oligo-FISH) technology. [Method] Based on the published cotton genome sequence, the chromosome-specific oligos library was obtained by bioinformatics method. Then, the obtained oligos were labeled through emulsion polymerase chain reaction, and used for FISH of cotton mitotic chromosome. [Results] A system of cotton Oligo-FISH technology was set up and successfully applied in cotton chromosome identification. [Conclusion] This system can be used for dissection of cotton genomes.

Keywords cotton      oligos      fluorescence in situ hybridization      chromosome     
Corresponding Authors: Fang Liu,Renhai Peng     E-mail: liuylay2012@163.com;liufcri@163.com;aydxprh@163.com
Issue Date: 02 June 2017
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Yuling Liu
Zhen Liu
Zhaoguo Li
Yuhong Wang
Zhongli Zhou
Xiaoyan Cai
Xingxing Wang
Xiaoyan Wang
Shulin Zhang
Haiyan Zhao
Zhenmei Zhang
Kunbo Wang
Fang Liu
Renhai Peng
Cite this article:   
Yuling Liu,Zhen Liu,Zhaoguo Li, et al. Construction and Primary Application of Oligos Fluorescence in situ Hybridization Technology in Cotton[J]. Cotton Science, 2017, 29(3): 213-221.
URL:  
http://journal.cricaas.com.cn/Jweb_mhxb/EN/10.11963/1002-7807.lylprh.20170427     OR     http://journal.cricaas.com.cn/Jweb_mhxb/EN/Y2017/V29/I3/213
Fig. 1  Procedure for design of single chromosome-specific oligos of cotton
水相Aqueous phase油相Oil phase
成分Component用量Usage /μL成分Component用量Usage /μL
Nuclease free water99.5矿物油Mineral oils10 000
5 × HF buffer30.0Triton X-1005
50 mmol L-1MgCl23.0ABIL EM90400
MYtags PCR Primer mix1.5
10 mmol L-1dNTP3.0
10 g L-1BSA (High Q)7.5
Table 1  Composition and quantity of aqueous phase and oil phase of emulsified PCR
Fig. 2  Detection of emulsion PCR product and labeled probe by gel
Fig. 3  Oligo-FISH on mitotic metaphase chromosomes ofGossypium raimondii
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