[1]李烨.茄子微卫星标记(SSR)反应体系的优化研究[J].北方园艺,2015,39(17):70-74.[doi:10.11937/bfyy.201517018]
　LI Ye.Optimization of SSR Analysis System in Eggplant[J].Northern Horticulture,2015,39(17):70-74.[doi:10.11937/bfyy.201517018]
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茄子微卫星标记(SSR)反应体系的优化研究

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第39卷 期数: 2015年17 页码: 70-74 栏目: 出版日期: 2015-09-15
Title:
Optimization of SSR Analysis System in Eggplant
作者:
李烨
（哈尔滨市农业科学院，黑龙江 哈尔滨 150070）
Author(s):
LI Ye
(Harbin Academy of Agricultural Science，Harbin,Heilongjiang 150070)
关键词:
茄子; 微卫星标记; 反应体系
Keywords:
eggplant; SSR; analysis system
DOI:
10.11937/bfyy.201517018
文献标志码:
A
摘要:
以茄子品种“哈农杂茄4号”DNA为试材，利用单因素试验方法，研究了茄子SSR技术中PCR体系的主要成分对SSR扩增结果的影响，并比较了聚丙烯酰胺凝胶和琼脂糖电泳检测扩增产物多态性的差异。结果表明:反应体系中，模板DNA的适宜浓度为20~40 ng，适宜的引物浓度为0.3 μmol/L，dNTP的适宜浓度为200 μmol/L，Mg2+的最适浓度范围为2.0~2.5 mmol/L，Taq聚合酶在20 μL反应体系中宜加入1 U。茄子SSR的最佳反应程序为94℃预变性5 min；94℃变性1 min，52~56℃复性1 min，72℃延伸2 min，35个循环;最后72℃延伸8 min。PCR产物检测时6%的聚丙烯酰胺凝胶的分辨率显著高于1.8%的琼脂糖电泳。
Abstract:
Taking eggplant DNA of ‘Hanongzaqie No.4’ as the test material，using single factor test method，the SSR amplified results of eggplant and comparative analysis of polymorphism of SSR detected on two gel electrophoresis systems were studied.A standard SSR system fitting for eggplant was established.The results showed that the optimized amplification conditions were 20-40 ng DNA，0.3 μmol/L of each primer，200 μmol/L of dNTP，2.0-2.5 mmol/L Mg2+ and 1 uint Taq DNA polymerse in 20 μL volume.The optimized PCR cycles started at 94℃(5 min)，followed by 35 cycles of 1 min at 94℃，1 min at 52-56℃ and 2 min at 72℃，and then extended 8 min at 72℃.6% Polyacrylamide gel had higher ability than 1.8% agarose gel in discriminating polymorphism.

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备注/Memo

作者简介：李烨(1972-)，女，硕士，研究员，研究方向为蔬菜遗传育种。E-mail：eggplant_2010@163.com.

更新日期/Last Update: 2015-10-10