TILLING (Targeting Induced Local Lesions in Genomes) is a process for producing a large number of recessive mutations by single-base substitution. This is in contrast to insertational mutagenesis or gene silencing methods. The process can produce several single base substitutions within one gene, the effects of which can be analysed experimentally.
In TILLING, mismatched strands of DNA with different alleles form bubbles called heteroduplexes, which are amplified by PCR. The mismatched DNA is cleaved by single stranded nucleases, and the products are separated by size. If there is a mutation, graphical analysis will show multiple peaks, corresponding to the main product and the snips from the heteroduplexes.
Mutants identified in TILLING may be incorporated into crop breeding programmes without using traditional GM technology.
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