KAPA血液DNA聚合酶扩增人全血基因组DNA的观察

Observing Study on the KAPA Blood DNA Polymerase Designed for Amplificating Human Whole Bood Gene

目的 验证KAPA血液DNA聚合酶扩增人全血基因组DNA的可行性.方法 运用12.5 μl KAPA全血PCR混合物分别扩增0.5,1.0,2.5和5 μl的EDTA全血中的白细胞介素-6基因启动子区一个片段,并与野生型聚合酶扩增效果进行比较.结果 琼脂糖凝胶电泳显示KAPA全血PCR试剂成功扩增出目的DNA片段,扩增产物浓度随EDTA全血体积的增大而升高.结论 KAPA全血PCR试剂可进行全血DNA的直接扩增,该方法具有简单、快速的特点,扩增产物可用于限制性内切酶切割,DNA测序或者dHPLC的分析.

Objective To validate the possibility of direct amplicating human gene from whole bood. Methods 12. 5μl of KAPA whole blood PCR mix and 0. 5,1.0,2.5,5μl of EDTA whole bood were used to amplify one fragment of interleukin-6 gene promoter area. The result was compared with wide -type DNA polymerase. Results Agarose gel electrophoresis result showed that target DNA fragment could be amplified by KAPA reagent. The concentration of amplication product increased with the volume of whole blood. Conclusion Whole blood DNA can be amplified directly by KAPA reagent. The method is simple and fast. PCR product can be used for the anlysis of RELP ,DNA sequening and dHPLC anlysis.