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Identifying Mutations by High Resolution Melting in a TILLING Population of Rice

Article DOI: 10.3791/59960-v • 06:10 min • September 2nd, 2019
September 2nd, 2019 •
1, 1, 1, 2, 3, 1, 4
1National Key Laboratory of Rice Biology, Zhejiang Provincial Key Laboratory of Crop Genetic Resources, Institute of Crop Science, Zhejiang University, 2Jiaxing Academy of Agricultural Sciences, 3Institute of Nuclear Agricultural Sciences, Zhejiang University, 4The New Countryside Development Institute at Zhejiang University

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In this article, we present the protocol that is described as high-resolution melting analysis (HRM)-based Target Induced Local Lesions In Genomes (TILLING). This method utilizes fluorescence changes during the melting of the DNA duplex and is suitable for high-throughput screening of both insertion/deletion (Indel) and single base substition (SBS).

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Mutations High Resolution Melting TILLING Population Rice Breeding Disease Resistance Mineral Content Cost-effective Technique 96-well PCR Plate Buffer A Solution Freezing Defreezing Incubation Buffer B Vortexing Centrifugation DNA Supernatant Annealing Temperature Optimization PCR Template Gradient-capable Thermal Block Electrophoresis Agarose Gels Amplicons Examination HRM Analysis Fluorescence Dye
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