TILLiNG (Targeting Induced Local Lesions in Genomes) is a reverse genetics technique designed to detect unknown mutations in a gene by forming DNA heteroduplex as a result of mixing wild-type and chemically-induced mutated genes.

In order to increase experimental throughput, the genes are pooled first, then PCR amplified. This is followed by heteroduplex formation, then the samples are enzymatically digested. Next, digested cleaved DNA fragments are analyzed for scoring. The enzyme specifically recognizes the mismatched base pairs and cleaves the DNA at the site of mismatch.

Until now, detecting unknown mutations was both a challenging and a time intensive process. Advanced Analytical has developed a streamlined TILLiNG process that requires NO fluorescently labeled primers and NO sample clean-up. Detecting unknown mutations has never been easier, especially with this process developed for the Fragment Analyzer™ Automated CE System platform (see below). In comparison to the traditional method, the Fragment Analyzer™ Automated CE System method requires less than ½ the hands on time and analyses can be completed twice as fast (total time) compared to competitive methods.

Currently, Fragment Analyzer™ Automated CE System customers are using their instruments to do:

• reverse genetics TILLiNG of rice.
• reverse genetics TILLiNG of corn.
• reverse genetics TILLiNG of wheat.
• reverse genetics TILLiNG of lettuce.
• reverse genetics TILLiNG of tomato.
• reverse genetics TILLiNG of soybean.

Regardless of whether your lab wants to do TILLiNG (mutation detection) with diploids, tetraploids or hexaploids, the Fragment Analyzer™ Automated CE System has excellent sensitivity for analyzing medium and large-sized TILLiNG pools.