| 香菇反转录转座子间扩增多态性（IRAP）PCR反应体系的研究 |
| |
| 引用本文： | 肖扬.香菇反转录转座子间扩增多态性（IRAP）PCR反应体系的研究[J].中国农学通报,2009,25(7):47-51. |
| |
| 作者姓名： | 肖扬 |
| |
| 作者单位： | 华中农业大学应用真菌研究所,武汉,430070 |
| |
| 基金项目： | 农业部公益性行业(农业)科研专项项目，农业部引进国际先进农业科学技术计划(948计划) |
| |
| 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 |
| 收稿时间： | 2008-12-08 |
| 修稿时间： | 2008-12-29 |
Study on PCR Reaction System of Inter-retrotransposon Amplified Polymorphism (IRAP) in Lentinula edodes |
| |
| XIAO Yang.Study on PCR Reaction System of Inter-retrotransposon Amplified Polymorphism (IRAP) in Lentinula edodes[J].Chinese Agricultural Science Bulletin,2009,25(7):47-51. |
| |
| Authors: | XIAO Yang |
| |
| Institution: | (The Institute of Applied Mycology, Huazhong Agricultural University, Wuhan 430070) |
| |
| 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. |
| |
| Keywords: | IRAP-PCR |
| 本文献已被 维普 万方数据 等数据库收录！ |
| 点击此处可从《中国农学通报》浏览原始摘要信息 |
| 点击此处可从《中国农学通报》下载免费的PDF全文 |
