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

Version 5 2016-05-04, 08:57
Version 4 2016-05-04, 08:55
Version 3 2016-05-04, 08:33
Version 2 2016-05-03, 09:35
Version 1 2016-05-03, 09:22
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posted on 2016-05-04, 08:57 authored by Jianxin 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.

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