遗传 ›› 2000, Vol. 22 ›› Issue (4): 197-200.

• 论文 •    下一篇

中间偃麦草单条染色体分离及体外扩增

李连城1;何聪芬1;2;徐琼芳1;马有志1;辛志勇1 LI Lian-cheng1;HE Cong-fen1;2;XU Qiong-fang1;MA You-zhi1;XIN-yong1   

  1. 1.中国农业科学院作物育种栽培研究所 农业部作物遗传育种重点开放实验室,北京 100081 2.河北师范大学 生物系, 石家庄 050016 1.Institute of Crop Breeding and Culture,CAAS,Bejing 100081,China 2.Biology Department of Hebei Normal University, Sijiazhuang 050016,Chian
  • 收稿日期:1900-01-01 出版日期:2000-08-10 发布日期:2000-08-10

Microisolation and Amplification in vitro of Single Th.intermedium Chromosome

  • Received:1900-01-01 Online:2000-08-10 Published:2000-08-10

摘要: 本研究利用微细玻璃针法从减数分裂中期I的花粉母细胞中分离回收了携带小麦抗黄矮病基因的中间偃麦草染色体2Ai-2。将目标染色体放入装有蛋白酶K消化液的0.5ml小离心管中,用Sau3AI酶切,在DNA片段的末端连接接头后,进行 PCR扩增。扩增产物经琼脂糖凝胶电泳分析,DNA片段的大小在150~3000bp之间,而大部分集中在200~1500bp之间。
Abstract:A simple method was used to adapt refined needles for the collection of Th.intermedium 2Ai-2 chromosome carrying BYDV (barley yellow dwarf virus) resistance gene from meiosis-metaphase spreads .The aimed chromosome was put into a 0.5ml Eppendorf tube,deproteinized with proteinase K,digested with Sau3AI,and link-adaptors were ligated to the ends of the DNA fragments.After amplification by PCR,size distributions of the PCR products were analyzed by agarose gel electrophoresis and smears of DNA were revealed that the size ranged from 150bp to 3000bp with predominant fragments at about 200~1500bp.

关键词: 染色体, 微分离, chromosome, 中间偃麦草
Key words,
LA-PCR