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Isothermal Amplification of Genetically Modified DNA Sequences Directly from Plant Tissues Lowers the Barriers to High-Throughput and Field-Based Genotyping
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2009
Year
EngineeringGeneticsDna AnalysisMolecular BiologyNucleic Acid Amplification TestMolecular GeneticsGenomicsPlant GenomicsWide Dynamic RangeBioanalysisTarget Dna SequencesMolecular DiagnosticsLoop-mediated Isothermal AmplificationDna SequencingField-based GenotypingDna ReplicationIsothermal AmplificationAgricultural BiotechnologyPlant Tissues LowersBiotechnologyGenetic EngineeringSynthetic BiologyNucleic Acid AmplificationMedicineGenetically Modified OrganismGenome Editing
DNA extractions are a major cost for high-throughput genotyping. The loop-mediated isothermal amplification (LAMP) assay has been used for the detection of two genetically modified (GM) related sequences. The amplification of target DNA sequences from leaf and maize seed tissues prepared with minimum preparative treatment (disruption in water) demonstrates the ability of LAMP to work in conditions normally inhibitive to PCRs. The wide dynamic range of detection in these samples suggests that LAMP is highly sensitive even when the target is presented in such a crude form. LAMP offers a means of reducing genotyping costs as well as simplifying testing procedures.