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- W4300100437 abstract "Target Induced Local Lesions In Genomes (TILLING) is a strategy of reverse genetics for the high-throughput screening of induced mutations. However, the TILLING system has less applicability for insertion/deletion (Indel) detection and traditional TILLING needs more complex steps, like CEL I nuclease digestion and gel electrophoresis. To improve the throughput and selection efficiency, and to make the screening of both Indels and single base substitions (SBSs) possible, a new high-resolution melting (HRM)-based TILLING system is developed. Here, we present a detailed HRM-TILLING protocol and show its application in mutation screening. This method can analyze the mutations of PCR amplicons by measuring the denaturation of double-stranded DNA at high temperatures. HRM analysis is directly performed post-PCR without additional processing. Moreover, a simple, safe and fast (SSF) DNA extraction method is integrated with HRM-TILLING to identify both Indels and SBSs. Its simplicity, robustness and high throughput make it potentially useful for mutation scanning in rice and other crops." @default.
- W4300100437 created "2022-10-03" @default.
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- W4300100437 date "2019-09-02" @default.
- W4300100437 modified "2023-09-26" @default.
- W4300100437 title "Identifying Mutations by High Resolution Melting in a TILLING Population of Rice" @default.
- W4300100437 doi "https://doi.org/10.3791/59960-v" @default.
- W4300100437 hasPublicationYear "2019" @default.
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