RACHITT

Coco, W.M. (2003) RACHITT gene family shuffling by random chimeragenesis on transient templates. Methods in Molecular Biology (Clifton, NJ), 231, 111-127. 

Coco, W. M., RACHITT Gene family shuffling by Random Chimeragenesis on Transient Templates. Methods. Mol.Biol., 2003. 231 pp. 111-127. 

A library of parent DNA sequences encoding for the desired protein is chosen. Sequence diversity is created or increased through a mutagenesis step, either by introduction of random point mutations through error-prone PCR or by recombination of DNA fragments such as DNA shuffling or RACHITT. 

L-shuffling similar to RACHITT but lower diversity library due to use of larger fragments Proteus Corp. 

The reduced background of unshuffled clones is a significant advantage with these techniques, with over 75% shuffled clones in most cases and 100% with RACHITT. In addition, the higher crossover frequency observed with RACHITT, and the average of 14 per gene compared to one to four for other PCR-based methods, leads to a considerable increase in library diversity (Pelletier, 2001). 

Figure 11.6 Random Chimeragenesis on Transient Templates (RACHITT) with mutagenesis (Gibbs, private communication).

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