蓖麻茎秆cDNA-AFLP反应体系的建立  

Establishment of cDNA-AFLP Analysis System for Castor

作　　者：王文跃[1,2] 李国瑞[1,2] 黄凤兰[1,2] 尚雨丝[1] 王超[1] 罗蕊[1] 邱靖[1] 王丹[3] 张智勇[3] 陈永胜[1,2] 

机构地区：[1]内蒙古民族大学,内蒙古通辽028000 [2]内蒙古自治区高校蓖麻产业工程研究中心,内蒙古通辽028000 [3]通辽市农业科学研究院,内蒙古通辽028000

出　　处：《华北农学报》2013年第2期86-90,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(30760123);国家自然科学基金项目(31160290);内蒙古自然科学基金(2009BS0304);内蒙古自然科学基金(2010BS0511);国家民委项目(10NM02)

摘　　要：以蓖麻茎秆为试材,研究了影响cDNA-AFLP标记蓖麻差异基因反应体系的几个关键因素,建立了适宜蓖麻的cDNA-AFLP分析体系。结果表明:当裂解液为1.4 mL时提取的RNA浓度最高,EcoRⅠ和MseⅠ的组合6 h可将ds-cDNA酶切完全,与相应接头连接过夜后产物用于预扩增,预扩增产物稀释15倍时作为选择性扩增模板可以获得条带丰富且重复性好的结果。蓖麻cDNA-AFLP反应体系的建立为进一步研究蓖麻株高、叶型等相关基因奠定了基础。In this paper,we studied the factors affecting cDNA-AFLP marked castor differential gene,established the suitable cDNA-AFLP analysis system for caster.The results indicated that the concentration of the RNA can be the highest when the lysate volume was 1.4 mL.The enzyme combination EcoR Ⅰ and Mse Ⅰ digested ds-cDNA thoroughly for 6 h.Then it was linked with the connection and the product was used for pre-amplified.The reproducible result and the rich bands can be obtained that the pre-amplified product was diluted 15-fold then was selective amplified.The research could provide important evidence for further studying the gene of caster on plant height,leaf type and so on.

关 键 词：CDNA-AFLP 蓖麻 体系建立 

分 类 号：S565.6[农业科学—作物学]