DMSO对梨属植物ITS区扩增及测序的影响

【作者】 郑小艳； 蔡丹英； 滕元文；

【机构】 浙江大学农业与生物技术学院园艺系；

【摘要】 普通PCR体系中加入4％的DMSO了改进了梨属植物(Pyrus)ITS(internal transcribed spacer)区的扩增和测序性能。结果表明适量的DMSO作为一种辅助剂不仅显著提高了扩增效率和特异性,而且有效的防止了不同长度的ITS假基因序列的扩增,使得PCR 产物直接测序得到理想的效果,为基于ITS序列进行梨属植物系统分析奠定了基础。更多还原

【Abstract】 The PCR amplification and sequencing of ribosomal ITS (internal transcribed spacer) region of Pyrus L. was improved by the addition of 4 % DMSO in basic PCR protocol. It was found that DMSO used as an adjuvant greatly increased the amplification efficiency and specificity of PCR, and different kinds of ITS pseudogene .sequences were prevented from being amplified. As a result, ideal direct sequences of PCR products were obtained, which will provide the basis for reconstructing a phylogenetic relationship of Pyrus.更多还原