| 7对猪源性成分检测相关引物的特异性评测 |
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| 引用本文： | 顾佳瑛,薛超波,宋蓉,林昕,管峰.7对猪源性成分检测相关引物的特异性评测[J].安徽农学通报,2015(17):121-124. |
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| 作者姓名： | 顾佳瑛 薛超波 宋蓉 林昕 管峰 |
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| 作者单位： | 1. 中国计量学院生命科学学院,浙江杭州,310018;2. 舟山市食品药品检验检测研究院,浙江舟山,316021 |
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| 基金项目： | 浙江省食品药品监督局项目(2014020)，中国计量学院实验室开放项目。 |
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| 摘 要： | 为了评测物种鉴定PCR体系的特异性及其影响因素,实验对猪源性成分鉴定相关的7对引物就退火温度、镁离子浓度和不同Taq酶的影响进行了研究。结果表明,退火温度和镁离子浓度对PCR特异性有着重要影响,而其中2对引物优化后仍与鼠、兔、牛、羊、鸡和鸭有非特异性扩增。不同的Taq酶对引物特异性也有重要影响,本实验中r Taq酶在7对引物PCR体系中特异性最佳。通过对引物体系的优化和选择不同的Taq酶,可以有效提高特异性,减少非特异性扩增,为建立物种鉴定的PCR体系及多重PCR提供了基础。
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| 关 键 词： | 猪 物种鉴定 引物 特异性评测 |
Assessment and Determination of Specificity of Seven Pairs of Primers for Porcine Identification |
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| Abstract: | To assess and determine the specificity of PCR system for species identification,seven pairs of primers for porcine identification were used to assess the effects of annealing temperature,concentration of MgCL2 and different Taq DNA polymerase on PCR specificity in this study. The results showed that there had two of seven pairs of primers had nonspecific amplification with rat,rabbit,bovine,ovine and chicken after optimization of PCR conditions. After PCR conditions were optimized for each pair of primers ,includ?ing annealing temperature,concentrations of MgCL2 and dNTP,different Taq DNA polymerases. The PCR specificity was improved but not significantly. The results indicated that rTaq polymerase enzyme performed best to ensure the specificity of seven pairs of primers. This study could provide some guidance for devel?opment multiplex PCR methods of species identification. |
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| Keywords: | Pig Species identification Primer Specificity assessment |
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