四种DNA聚合酶扩增效果的比较

Comparison of four DNA polymerases for amplication effects

目的比较及评价不同品牌的DNA聚合酶的扩增效果,为筛选性价比高的酶提供参考。方法本实验选用四个品牌的DNA聚合酶,设定不同的模板浓度、产物长度、退火温度等进行PCR扩增,扩增产物通过琼脂糖凝胶电泳检测扩增效果。结果四个品牌的聚合酶通过灵敏度检测的检测下限都在10 pg/μl以下,退火温度在55~65℃范围都能扩增出目的条带,对退火温度的适应范围比较广。品牌4扩增出的目的条带最多,扩增范围最广;品牌2电泳条带最明亮,扩增能力最强;品牌1条带最单一,扩增特异性最好。结论四个品牌的酶在灵敏度、扩增长度、退火温度范围、扩增时间等方面都有一定差异。品牌2扩增能力最强,性价比最高,更适合工业领域。

Objective To compare and evaluate the amplification effect of DNA polymerase from different manufacturers to guide screen DNA Polymerases with high cost performance.Methods In this experiment,DNA polymerases from four brands were used for PCR amplification with different template concentration,product length and annealing temperature.The amplification effect was detected by agarose gel electrophoresis.Results The detection limit of four brands were below 10 pg/μl.The annealing temperature can be amplified in the range of 55-65℃,and the adaptation range of the four brands of enzymes to the annealing temperature is relatively wide.Brand 4 expands the most destination bands and the widest expansion range.The DNA polymerases amplification ability of brands 2 was the strongest,and the electrophoresis band of amplification result was the brightest.The DNA polymerases amplification specificity of brands 1 was the best,and the electrophoresis band of amplificationresult was the most single.Conclusion The DNA polymerases of the four brands have certaindifferences in sensitivity,annealing temperature range,amplificationlength and amplification time.DNA polymerase of brands 2 has the strongest amplification ability and the highest cost performance,which is more suitable for industrial field.