Effects of different end primer bases on PCR amplification product

ObjectiveTo explore the effects of different bases at 3' end of the primer on products of polymerase chain reaction.MethodsHBVg ene was amplified by PCR in the same forward primer and anneal temperature.Recombinan plasmid pUC-19 carrying whole HBVgenome was used as a template and the base at the 3' end of the reverse primer was T (AP1),G(AP2),A(AP3),and C(AP4),respectively.ResultsThe form of production and primer dimer of different reaction systems in the same anneal temperature were different.When anneal temperature was 50℃,reaction systems of AP1 and AP4 had visible amplification products.AP3 had small quantities of amplification products.Amplification products of AP2 could not be observed and non-specific products were generated.When anneal temperature was 55℃,reaction systems of AP1 and AP2 had visible amplification products;AP4 had small quantities of amplification products.Amplification products of AP2 could not be observed.Reaction systems of AP1 and AP4 had the same quantities of primer dimers.Reaction systems of AP2 and AP3 had less prmier dimers.When anneal temperature rised to 62℃,reaction systems of AP1 and AP2 had visible amplification products and AP4 had small quantities of amplification products.Amplification products of AP3 could not be observed,while primer dimers coulde be observed.ConclusionThe different bases at 3' end of the primer effectes deeply on products of PCR.