TIAN Chang-ping,ZHANG Fu-xing,LIU Xiao-jing,et al.Optimization and Application of Fluorescent-AFLP Analysis System on Cherry[J].Northern Horticulture,2014,38(14):115-118.
樱桃荧光AFLP反应体系优化及应用
- Title:
- Optimization and Application of Fluorescent-AFLP Analysis System on Cherry
- 文章编号:
- 1001-0009(2014)14-0115-04
- Keywords:
- cherry; fluorescent-AFLP; optimization; application
- 分类号:
- S 662.5
- 文献标志码:
- A
- 摘要:
- 以12个甜樱桃品种和4个中国樱桃品种为试材，对荧光AFLP分析过程中模板DNA的制备、酶切与连接、酶切连接产物的预扩增与选择性扩增、聚丙烯酰胺凝胶电泳等过程中易出现的问题及其解决方法进行了研究。结果表明：改良CTAB法提取甜樱桃嫩叶和老叶DNA效果均很好；200 ng DNA 双酶切反应4 h，连接产物稀释10倍作为预扩增模板，预扩增产物稀释20倍作为选择性扩增的模板；筛选了64对引物，其中8对扩增效果理想，能够得到清晰、稳定的条带，平均每对引物扩增条带为40.6条，建立了樱桃AFLP分子标记技术体系；该体系的建立为研究甜樱桃遗传多样性和标记重要农艺性状奠定了基础。
- Abstract:
- Taking 12 sweet cherry cultivars and 4 Chinese cherry cultivars as materials for establishing DNA-AFLP system.Several key factors affecting DNA-AFLP analysis such as DNA extraction,enzymes restriction,adaptor-ligation,pre-amplification and selective amplification were detected.The results indicated that CTAB method could be used for extracting ideal DNA from leaf of sweet cherry;200 ng DNA was digested completely by EcoR I and Mse I for 4 h.ligation products were diluted to 10 times for pre-amplification template and pre-amplification products were diluted to 20 times for selective amplification-64 pairs of primers were screened,of which 8 pairs could amplify clear,stable and satisfactory results an average of each primer band was 40.6.This study established a set of AFLP marker system for cherry,providing a foundation for studying genetic diversity and important agronomic traits.
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备注/Memo
第一作者简介：田长平（1983-），男，农艺师，现主要从事樱桃栽培与育种等研究工作。E-mail:wenjun8341@163.com.