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《Journal of Anhui Agricultural Sciences》 2009-18
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An Efficient Method of Nuclei Extraction in Cotton

PENG Ren-hai et al(Key Laboratory of Cotton Genetic Improvement,Ministry of Agriculture/ Cotton Research Institute,Chinese Academy of Agricultural Sciences,Anyang,Henan 455000)  
[Objective] The experiment aimed to study an efficient method of nuclei extraction of cotton and provided technical support for constructing large-insert genomic library and sequencing complete genome.[Method] The cotton cotyledons germinated in dark moisture chamber for one week were chopped with a sharp sterile scalpel in a Petri dish which contained ice-cold nucleus isolation buffer(10 mmol/L MgSO4,5 mmol/L KCl,0.5 mmol/L HEPES,1 mg/ml DTT,0.25% Triton X-100 and 2% PVP40),then the nuclei were collected after selected through 100,50 and 30 μm nylon meshes and centrifugation.Meanwhile,the tender leaves and cotyledons with different germination time in dark were treated by grinding method and Jackknife method.[Result] The chopping with a sharp scalpel method was very simple and rapid,which did not need grind and the success rate was 100%.[Conclusion] An efficient method of nuclei extraction of cotton with simple,high efficiency,rapid reaction and poison free were established.
【Key Words】： Cotton Nuclei Chopping with a sharp scalpel
【Fund】： 国家自然科学基金(30170501);; 国家转基因生物新品种培育重大专项(2008ZX08005-003);; 国家高技术研究发展计划(2003AA207051)
【CateGory Index】： S562
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