CAJ | 학술논문

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。
위료평측물충감정PCR체계적특이성급기영향인소，실험대저원성성분감정상관적7대인물취퇴화온도、미리자농도화불동Taq매적영향진행료연구。결과표명，퇴화온도화미리자농도대PCR특이성유착중요영향，이기중2대인물우화후잉여서、토、우、양、계화압유비특이성확증。불동적Taq매대인물특이성야유중요영향，본실험중r Taq매재7대인물PCR체계중특이성최가。통과대인물체계적우화화선택불동적Taq매，가이유효제고특이성，감소비특이성확증，위건립물충감정적PCR체계급다중PCR제공료기출。
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.