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posted on 2016-05-04, 08:57authored byJianxin YangJianxin Yang, Berit Kemps-Mols, Marijke Spruyt-Gerritse, Jacqueline Anholts, Frans Claas, Michael Eikmans
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.