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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (7): 47-51.

所属专题： 生物技术； 食用菌； 食用菌

• 生物技术科学 • 上一篇    下一篇

香菇反转录转座子间扩增多态性（IRAP）PCR反应体系的研究

肖扬   

  • 收稿日期:2008-12-08 修回日期:2008-12-29 出版日期:2009-04-05 发布日期:2009-04-05

Study on PCR Reaction System of Inter-retrotransposon Amplified Polymorphism (IRAP) in Lentinula edodes

XIAO Yang   

  • Received:2008-12-08 Revised:2008-12-29 Online:2009-04-05 Published:2009-04-05

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摘要： 摘 要：为了开发香菇反转录转座子间扩增多态性标记(IRAP)，建立稳定的IRAP-PCR反应体系，本文对影响IRAP-PCR的主要因素进行了优化筛选。确定了最佳反应体系为：20μl反应体系中，包含30 ng模板DNA，0.30μmol/L引物， 0.3mmol/L dNTPs，2.0 mmol/L Mg2+及0.75UTaq酶。梯度PCR试验筛选得到的最佳退火温度为56.1℃。采用上述最佳反应体系和引物LTR1L-MarY1L对香菇18个菌株进行了IRAP-PCR扩增，验证了该体系的可靠性。

关键词: 覆盖方式, 覆盖方式, 烤烟, 叶绿素, 酶活性, 丙二醛

Abstract: Abstract:. The main factors affecting IRAP-PCR amplification were optimized in order to develop inter-retrotransposon amplified polymorphism marker and to establish stable IRAP reaction system in Lentinula edodes. The optimal IRAP-PCR system was as follows, within 20μl reaction system, there were 30ng template DNA, 0.3μmol/L primer, 0.3mmol/L dNTPs, 2.0mmol/L Mg2+ and 0.75U Taq DNA polymerase. The optimal annealing temperature was determined as 56.1℃ by gradient PCR. Based on the optimal IRAP-PCR system, primer combination LTR1L and MarY1L was used to amplify genomic DNA of 18 strains. Results indicated that the optimal IRAP-PCR system is reliable.