摘要
以改良的苜蓿雄性不育系叶片为材料,通过RNA提取、反转录、酶切、链接、PCR扩增、凝胶电泳等系列程序的摸索和优化,建立了适宜苜蓿cDNA-AFLP的反应体系,并得到了清晰可辨的cDNA-AFLP指纹图谱。结果如下:cDNA经EcoRⅠ和MseⅠ完全酶切后,16℃连接15 h;20μl预扩增体系中加入连接产物2.0μl较适宜,预扩增产物稀释20倍后进行选择性扩增效果最佳。
cDNA - AFLP reaction system for alfalfa was established with the leaves of modified male sterile line, and clear fingerprint spectrum were obtained. A series of procedures were the RNA extraction, reverse transcription, digestion, links, PCR amplification and gel electrophoresis. The results were as follows ：cDNA was linked under 16 ℃ for 15 h after it was digested with EcoR Ⅰ and Mse Ⅰenzyme. It was suitable that took Connect product 2.0μl into the 20μl pre-amplification System for pre-amplification. The effect was the best that the pre-amplified product was diluted 20-fold then was selective amplified.
出处
《种子》
CSCD
北大核心
2011年第8期1-4,8,共5页
Seed
基金
国家自然科学基金(31060321)
内蒙古自然科学基金(2010Zd22)
内蒙古农村领域科技计划项目(20091401)
中央级公益性科研院所基本科研业务费专项(牧草种质资源收集
评价及创新研究)
内蒙古社会发展领域科技计划项目(20082002)
