Some U.S bioethanol plants use a blend of maize and sorghum grains as feedstocks depending on availability and price. Knowledge of the grain composition of milled feedstock is beneficial for maximizing process efficiency and for potential future regulatory requirements. To analyze feedstock composition, we developed a duplex real-time PCR assay based on TaqMan\u2122 probes for the simultaneous analysis of flour blends. For sorghum, the method amplified a sequence within the F-Box domain-containing protein and for maize, the method amplified a sequence of the high mobility group a (HMGa) proteins. To acquire fluorescent signals simultaneously, FAM and TAMRA fluorescent probes were used to label the sorghum- and maize- specific probes, respectively. Results showed that the primers and probes were highly specific either for maize or sorghum. The real-time duplex PCR assay was validated on simulated flour binary mixtures and high in the precision and accuracy for the sorghum PCR system. Results with the established method showed specificity and sensitivity in quantifying sorghum composition in bioethanol feedstock mixtures.
\nResources in this dataset:
\nResource title: Primer sets used\nFile name: Table 1 1.jpg
\nResource title: Experimental setup\nFile name: experiemntal set ups Real time PCR project Sorgh-Maize.xlsx
\nResource title: Multiplex data HMGa and FBox\nFile name: 2022-01-24 multiplex FBox - HMGa 60 C 200nM.xls\nResource description: Data and analysis of sorhum maize and miusxtures using HMGa and FBox genes
\nResource title: multiplex FBox HMGa flour mixes 60 C\nFile name: 2022-02-22 multiplex FBox HMGa flour mixes 60 C 11 points.xls\nResource description: PCR multiplex data FBox HMGa flour mixes 60 C
\nResource title: Standard curves for zein DNA HMGa gene\nFile name: 2022-01-11 Hmga - Zein X07535 standard curves.xls\nResource description: Standard curves for zein DNA HMGa gene detection
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