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1.4mL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。
王文跃
,
李国瑞
,
黄凤兰
,
尚雨丝
,
王超
,
罗蕊
,
邱靖
,
王丹
,
张智勇
,
陈永胜
. 蓖麻茎秆cDNA-AFLP反应体系的建立[J]. 华北农学报, 2013
, 28(2)
: 86
-90
.
DOI: 10.3969/j.issn.1000-7091.2013.02.016
In this paper,we studied the factors affecting cDNA-AFLP marked castor differentialgene,estab-lished 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. 4mL. 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 re-producible result and the rich bands can be obtained that the pre-amplified product was diluted 15-fold then was se-lective amplified. The research could provide important evidence for further studying thegene of caster on plant height,leaf type and so on.
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