The source of SYBR Green master mix determines outcome of nucleic acid amplification reactions

Numerous real-time PCR devices and master mixes are available on the market. To perform reliable high-quality data, PCR master mix, and equipment need to be optimal. However, general lab optimized protocols are widely used for different gene targets and performed diversely between conditions. Our main objective was to test the robustness of different commercial SYBR green PCR mixes with respect to specificity and sensitivity of the PCR assay. This was tested across various PCR machines and amplification protocols for assessment of mRNA transcript levels, DNA copy numbers, and DNA genotypes.

目前市场上存在大量实时荧光定量PCR（real-time PCR）仪器与PCR预混试剂（PCR master mix）。为获取可靠且高质量的实验数据，所使用的PCR预混试剂与仪器需处于最优状态。然而，当前多数实验室的优化实验方案被广泛应用于不同基因靶点的检测，且不同实验条件下的操作流程存在显著差异。本研究的核心目标为：针对PCR检测的特异性与灵敏度，评估不同商用SYBR Green PCR预混试剂（SYBR green PCR mixes）的鲁棒性。本次评估覆盖多款PCR仪器与多种扩增方案，用于mRNA转录水平、DNA拷贝数及DNA基因型的检测。