[1]聂兴华,张卿,刘阳,等.板栗SSR标记研究中聚丙烯酰胺凝胶银染显影体系的优化[J].北方园艺,2019,43(12):27-33.[doi:10.11937/bfyy.20180369]
　NIE Xinghua,ZHANG Qing,LIU Yang,et al.Optimization of PAGE Silver-stained System Based on SSR Markers in the Chestnut[J].Northern Horticulture,2019,43(12):27-33.[doi:10.11937/bfyy.20180369]

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板栗SSR标记研究中聚丙烯酰胺凝胶银染显影体系的优化

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 43卷期数: 2019年12 页码: 27-33 栏目: 出版日期: 2019-06-30

Title:: Optimization of PAGE Silver-stained System Based on SSR Markers in the Chestnut

作者:: 聂兴华1; 张卿1; 2; 刘阳1; 代月星1; 曹庆芹3; 秦岭1; （1.北京农学院植物科学技术学院，北京 102206；2.农业应用新技术北京市重点实验室，北京 102206；3.北京农学院生物科学与工程学院,北京 102206）

Author(s):: NIE Xinghua1; ZHANG Qing1; 2; LIU Yang1; DAI Yuexing1; CAO Qingqin3; QIN Ling1; (1.College of Plant Science and Technology,Beijing University of Agriculture,Beijing 102206;2.Beijing Key Laboratory for Agricultural Application and New Technique,Beijing 102206;3.College of Biological Science and Engineering,Beijing University of Agriculture，Beijing 102206)

关键词:: 聚丙烯酰胺凝胶; 板栗; 正交实验; 银染显影效果

Keywords:: polyacrylamide gel; chestnut; orthogonal experiment; silver-stained effect

DOI:: 10.11937/bfyy.20180369

文献标志码:: A

摘要:: 为筛选出高效、稳定的银染显影体系，以板栗叶片DNA为扩增模板，利用L16(45)正交实验设计的方法，研究了AgNO3使用量、CH2O使用量、Na2CO3使用量及上样量4个主要因素对聚丙烯酰胺凝胶银染显影的影响，并对优化出的方法在银染效果、试剂使用和显影时耗等方面进行分析。结果表明：在碱性条件下，0.1% AgNO3+1 mL CH2O+0.2 g Na2CO3（500 mL体系）+1.5 μL上样量的银染显影效果最佳。筛选出的体系能有效克服胶图分辨率低、背景暗，读带不清晰和危险试剂消耗大的问题，为批量试验、快速读带提供了支持。

Abstract:: In order to screen out the high-efficiency and stable silver-stained system，in the study the DNA of chestnut leaves were used as the amplification template,and the four levels of AgNO3 usage amount,CH2O usage amount,Na2CO3 usage amount and sample volume were filtrated by L16(45) orthogonal experimental design,which were proved how these four main factors affect the polyacrylamide gel silver-stained effect.The improved method was analyzed in terms of silver-stained effect，reagent and silver-stained time.The results showed that under alkaline conditions，0.1% AgNO3+1 mL CH2O+0.2 g Na2CO3 (500 mL system)+1.5 μL samplevolume was the best.The selected system could effectively overcome the issues of low resolution，dark background，unclear read bands and high consumption of dangerous reagents.It provided support for batch experiments and fast reading.

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备注/Memo

第一作者简介：聂兴华(1992-)，男，硕士研究生，研究方向为果树发育生物学。E-mail：niexinghuabua@163.com.责任作者：张卿（1979-），男，博士，实验师，研究方向为实验技术和果树种质资源学。E-mail：zhangqing@bua.edu.cn.基金项目：河北科技计划专项资助项目（162268230）；北京市属高等学校创新团队建设与教师职业发展计划资助项目（IDHT20180509）；北京市自然科学基金重点（B）类资助项目（KZ201710020012）。收稿日期：2018-12-13

更新日期/Last Update: 2019-07-17