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The FF allele construct was generated from the VDRff plasmid by site directed mutagenesis by utilizing the QuickChange site directed mutagenesis kit (Stratagene, La Jolla, CA).

In summary, screening for genes important for mitochondrial function by utilizing gene trap mutagenesis and FACS in mouse ES cells is an effective approach that offers the potential to generate novel animal models and to identify genes that might not be identified from screens in other cell types.

By utilizing a transposon-based mutagenesis approach, we isolated a swarm-defective mutant of S. typhimurium (21).

The Brachyury response element in the p21 promoter was mutated by utilizing the GENEART site-directed mutagenesis system (Invitrogen Corp).

By utilizing the approach of signature tagged mutagenesis (STM) [32] [34], colony morphology variants were identified by screening a STM minitransposon library grown under different culture conditions in vitro.

Using reverse engineering and site directed mutagenesis methods, we were able to confirm that the amino acid substitution of the Asp285 is responsible for the increased aldehyde reductase activities by utilizing the additional cofactor NADPH.

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The mutated version of this plasmid (p-zbtb7a 3'UTRm) was generated by utilizing the p-zbtb7a 3'UTR as template and modifying the miR-20a seed binding site using the QuikChange II XL Site-Directed Mutagenesis Kit.

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Primers (Table 1 of the Supporting Information) were designed, and site-directed mutagenesis was performed according to the QuikChange mutagenesis protocol (Agilent) utilizing Pfu DNA polymerase.

PIPE mutagenesis utilizing overlapping complementary 5′ primer ends was used to amplify mutant constructs for all mutants.