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巢蕨基因组DNA提取与RAPD反应体系优化
引用本文:王雪兵,于旭东,朱会军,吴繁花,黄小凤,胡新文.巢蕨基因组DNA提取与RAPD反应体系优化[J].热带生物学报,2012,3(4):379-383.
作者姓名:王雪兵  于旭东  朱会军  吴繁花  黄小凤  胡新文
作者单位:1. 海南大学农学院,海南海口570228;东莞植物园,广东东莞523086
2. 海南大学农学院,海南海口570228;海南大学热带作物种质资源保护与开发利用教育部重点实验室,海南,海口570228
3. 海南大学农学院,海南海口570228;贵州大学精细化工研究开发中心/绿色农药与农业生物工程国家重点实验室培育基地/教育部重点实验室,贵州贵阳550000
4. 海南大学农学院,海南海口,570228
5. 东莞植物园,广东东莞,523086
基金项目:国家自然科学基金项目(31060123,30760025);海南省自然科学基金项目(311021,312056);海南大学教育教学研究课题立项项目(hdjy0920);东莞市高等院校、科研机构科技计划项目(2011108102039,2012108102054)
摘    要:以新鲜幼嫩的巢蕨叶片为材料,优化了巢蕨叶片基因组DNA的提取方法与RAPD反应体系.结果表明,利用改进的CTAB方法,能够提取出高质量的巢蕨基因组DNA;优化的巢蕨RAPD的最终反应体系(25μL)为:模板DNA(50 mg·L-1)1.5 μL,引物(1 mmol·L-) 1.5 μL,2×EasyTaq PCR SuperMiX 12.5 μL.

关 键 词:巢蕨Neottopteris  nidus  DNA提取  RAPD  分子标记

Extraction of Genomic DNA of Neottopteris nidus and Optimization of the RAPD Reaction System
WANG Xue-bing,YU Xu-dong,ZHU Hui-jun,WU Fan-hu,HUANG Xiao-feng and HU Xin-wen.Extraction of Genomic DNA of Neottopteris nidus and Optimization of the RAPD Reaction System[J].Journal of Tropical Biology,2012,3(4):379-383.
Authors:WANG Xue-bing  YU Xu-dong  ZHU Hui-jun  WU Fan-hu  HUANG Xiao-feng and HU Xin-wen
Institution:1. College of Agronomy, Hainan University, Haikou 570228, China; 2. Research and Development Center for Fine Chemicals/State Key Laboratory Breeding Base for Green Pesticide and Agricultural Bioengineering/ Ministry of Education Key Laboratory,Guiyang, Guizhou 550000, China; 3. Ministry of Education Key Laboratory for Conservation and Utilization of Tropical Crops Germplasm Resources, Haikou 570228, China; 4. Dongguan Botanic Garden, Dongguan, Guangdong 523086, China)
Abstract:In order to study the genitic diversity and genetic structure of Bird' s nest fern ( Neottopteris nidus ) populations, fresh tender fronds of the Bird' s nest fern were collected and resorted to genomic DNA extraction through optimized RAPD reaction system by using different concentrations of CTAB and NaC1, different volumes of template DNA and primer. The improved CTAB DNA extraction method was the best to extract the genomic DNA of the fronds of the Bird's nest fern. The optimum reaction system (25 μL) for RAPD amplification of the Bird' s nest fern DNA was listed as follows : The template DNA (50 mg · L- 1 ) 1.5 μL; the primer ( 1 mmol ·L-1) 1.5 μL; 2 ×EasyTaq PCR SuperMiX 12.5 μL.
Keywords:Bird's nest fern  Neottopteris nidus  Asplenium nidus  extration of genomic DNA  RAPD  molecular marker
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