一种利用双分子标记鉴定蓖麻种子品种真实性的方法及试剂盒

Method and kit for authenticating variety of castor seed by means of dimolecular markers

Abstract

本发明涉及一种利用双分子标记鉴定蓖麻种子品种真实性的方法及试剂盒,步骤如下:(1)提取蓖麻种子基因组DNA:(2)以步骤(1)获得的基因组DNA为模板进行AP-PCR扩增,引物见序列1;(3)对步骤(2)获得的AP-PCR扩增产物进行凝胶电泳;(4)以步骤(1)获得的基因组DNA为模板进行RMAPD扩增,引物见序列2和序列3;(5)对步骤(4)获得的RMAPD扩增进行聚丙烯酰胺凝胶电泳;(6)将未知品种的条带与已知品种数据库进行比对,判断蓖麻种子的品种。本方法采用自主开发的试剂盒和实验室常规的PCR技术与电泳技术,从取样到检测完成只需7h左右,且周年都可以进行。简单易行、快速灵敏、结果准确。
The invention relates to a method and kit for authenticating the variety of a castor seed by means of dimolecular markers. The method comprises the following steps: (1) extracting the genome DNA (deoxyribonucleic acid) of the castor seed; (2) using the genome DNA obtained in the step (1) as the template to carry out AP-PCR (arbitrarily primed polymerase chain reaction) amplification, wherein the primer is shown in sequence 1; (3) carrying out gel electrophoresis on the AP-PCR amplification product obtained in the step (2); (4) using the genome DNA obtained in the step (1) as the template to carry out RMAPD (random microsatellite amplification polymorphic DNA) amplification, wherein the primers are shown in sequence 2 and sequence 3; (5) carrying out polyacrylamide gel electrophoresis on the RMAPD amplification product obtained in the step (4); and (6) comparing the bands of the unknown variety with a data base of known varieties to judge the variety of the castor seed. By using the self-developed kit and the laboratory conventional PCR (polymerase chain reaction) technology and electrophoretic technology, the whole process from sampling to detection can be finished in about 7 hours, and the method can be implemented throughout the whole year. Besides, the invention has the advantages of high simplicity and practicability, high speed, high sensitivity and accurate result.

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NO-Patent Citations (2)

    Title
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