Utilization of corn silk for GMO detection through real time PCR

Abstract Purpose: Corn silk is a fiber rich byproduct obtained during corn processing. Present investigation deals with the purpose of GMO detection and quantification and comparison of DNA extraction in corn silk that is considered generally as a waste product. Method: A comprehensive analysis of extracted DNA quality, extraction cost, and time and PCR amplification was performed. Samples were homogenized by three different methods i.e. liquid nitrogen (CSK-LN), tissue lyzer (CSK-TL) and by grinder (CSK-RG). Afterwards, DNA extraction and quantitative analysis of extracted DNA was done using three different Kits QIAGEN DNeasy Mericon Food Kit, eurofins|GeneScan GENESpin DNA isolation Kit and PENICON EZ-10 Spin Column Genomic DNA Miniprep Kit .Furthermore, the agarose gel electrophoresis assay of corn silk was performed. Also scanning electron microscopy revealed observable differences between all 3 samples.Results: CSK-TL showed finest appearance due to the nature of treatment received. Among all kits, kit 3 (PENICON EZ-10 Spin Column Genomic DNA Miniprep Kit) displayed the most economical one for extraction, however in terms of DNA quantification kit 1 (QIAGEN DNeasy Mericon Food Kit) showed highest % yield of 122.4 ng/ul followed by tissue lyzer technique. In the result of gel electrophoresis assay, 1000bp plus band size observed with different consistency according to different kits.Conclusion: DNA extracted using waste material (corn silk) can be used for GMO analysis through Real-Time PCR technique. Among different grinding techniques compared, Tissue lyzed sample (CSK-TL) displayed highest quantified DNA extracted. Hence, provided techniques could be helpful in determining DNA in waste products.