胍盐裂解液对自动化提取工作站DNA回收率的影响

Study on the effect of DNA recovery efficiency with guanidine salt solution in automation workstation

目的研究不同稀释度胍盐裂解液对自动化提取工作站DNA回收效率的影响。方法制备不同浓度的DNA样本,并将EQ1000法医DNA提取试剂盒中的胍盐裂解液梯度稀释,在本实验室配置的自动化提取工作站上,运行"工作站磁珠法"提取程序,AB-7500型荧光定量PCR仪检测提取前后的样本DNA浓度。选取2～7号梯度组DNA回收效率数据进行统计学检验,比较不同稀释度胍盐裂解液的处理对样品DNA回收效率的影响。结果 90%～60%稀释度胍盐裂解液组样品回收效率在66%左右;50%～20%稀释度组样品DNA回收效率在41%左右;稀释度在50%以下与60%以上组样品回收效率之间差异极显著。结论应用"工作站磁珠法"提取生物检材DNA,其DNA回收效率受胍盐裂解液浓度的影响。

Objective To study on the effect of DNA recovery efficiency with different concentration of guanidine salt solution in automation workstation.Methods DNA samples were prepared with different concentration,and the guanidine salt solution from the EQ1000 DNA extraction agent set were also diluted serially,and then they were used to DNA extraction with automation extraction workstation accoding to the＂workstation-magnetic bead＂ program.Subsequently,the concentration of DNA samples before and after extraction were detected by using the 7500 Real-Time PCR System.We choosed the DNA groups from No.2 to No.7 to estimating the effect of DNA recovery efficiency with different concentration of guanidine salt solution.Results About 66% of the recovery efficiency was observed for those groups extracted with 90%～60% dilution solution of guanidine salt,about 41% of the recovery efficiency with 50%～20% dilution solution of guanidine salt,and there was a significant difference for the DNA recovery efficiency between under 50% and above 60% groups.Conclusion When the biological sample DNA is extracted by ＂workstation-magnetic bead＂ method,the DNA recovery efficiency will be influenced by concentration of guanidine salt solution.