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作物学报 ›› 2017, Vol. 43 ›› Issue (08): 1139-1148.doi: 10.3724/SP.J.1006.2017.01139

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

玉米InDel分子标记20重PCR检测体系的建立

冯博1,2,**,许理文1,**,王凤格1,*,薛宁宁1,刘文彬1,2,易红梅1   

  1. 1北京市农林科学院玉米研究中心 / 玉米DNA指纹及分子育种北京市重点实验室,北京100097;2吉林农业大学农学院,吉林长春130118;3江苏省农业科学院农业生物技术研究所,江苏南京210014;4全国农业技术推广服务中心,北京100125
  • 收稿日期:2016-12-11 修回日期:2017-04-20 出版日期:2017-08-12 发布日期:2017-04-27
  • 通讯作者: 王凤格,E-mail: gege0106@163.com* E-mail:fengbo02220108@163.com
  • 基金资助:

    本研究由科技部国家科技支撑计划(2015BAD02B02)和北京市农林科学院院科技创新能力建设专项(KJCX20161501)资助。

Establishment of 20 PCR DetectionSystem with InDel Molecular Markers in Maize

FENG Bo1,2,**,XU Li-Wen1,**,WANG Feng-Ge1,*,XUE Ning-Ning1,LIU Wen-Bin1,2,YI Hong-Mei1,TIAN Hong-Li1,LV Yuan-Da3,ZHAO Han3,JIN Shi-Qiao4,ZHANG Li-Ke4,YU Rong-Hai2,ZHAO Jiu-Ran1   

  1. 1 Maize Research Center, Beijing Academy of Agriculture & Forestry Sciences / Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Beijing 100097, China; 2 College of Agriculture, Jilin Agricultural University, Changchun 130118, China; 3 Provincial Key Laboratory of Agrobiology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 4 National Agricultural Technical Extension and Service Center, Beijing 100125, China
  • Received:2016-12-11 Revised:2017-04-20 Online:2017-08-12 Published:2017-04-27
  • Contact: 王凤格,E-mail: gege0106@163.com* E-mail:fengbo02220108@163.com
  • Supported by:

    This research was supported by Grants from National Science and Technology Project from the Ministry,of Science and Technology of China (2015BAD02B02), and Science and Technology Innovation Special Project from Beijing Academy of Agriculture and Forestry Sciences (KJCX20161501).

摘要:

为构建玉米多重PCR检测体系,提高分子标记检测效率,利用10份代表性玉米材料对238对InDel引物进行单重PCR评估,共得到192对扩增效率高、稳定性好的引物。根据软件评估结果、扩增质量、产物范围、染色体均匀分布原则从192对引物中优选出30对综合表现较好的引物形成扩增产物范围在80~200 bp和200~400 bp的两组核心引物组合,每套组合中有10对引物分布在不同染色体上。在核心引物组合的基础上综合考虑染色体分布、碱基片段范围、引物荧光颜色,逐一添加引物,最终形成两组玉米20重PCR体系,一组40重荧光标记毛细管电泳。

关键词: 玉米, InDel, 多重PCR

Abstract:

In order to improve the detection efficiency with molecular marker , the multiple PCR detection system was constructed. In this study, 10 major materials were used to evaluate the single pair PCR with 238 pairs of InDel primers. According to the software quality evaluation results, product range, and the principle of chromosome uniform distribution, 30 pairs of primers were selected from 192 primers with better performance to form two groups of core primer combinations with amplified products in the range of 80-200 bp and 200-400 bp, There were10 pairs of primers distributing in different chromosomes for each primer combination. Based on core primer combination and comprehensive consideration on chromosome distribution, base fragment, and primers fluorescent color, we established two groups of corn test 20 PCR system and a group of 40 fluorescent capillary electrophoresis.

Key words: Maize, InDel, Multiplex PCR

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