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Later, deletion clones were generated using sub-cloning strategies involving PCR and restriction digestions (Fig. 1C).

Following manufacturer's instructions, clones were generated by a TOPO® TA Cloning kit (Invitrogen, Carlsbad, CA).

Deletion and point substitution mutant clones were generated using a QuikChange Site-Directed Mutagenesis Kit (Agilent, Clara, CA).

A total of 239 unique clones were generated from 1447 recombinants, and 214 unique sequences were obtained.

McBride et al. (2008) founded populations of viruses from the clones with high, respectively low robustness (how these clones were generated will be described in a later section).

Individual clones were generated by limited dilution and propagated further.

Clones were generated using the MARCM method [32].

ept mutant eye clones were generated as described previously [4], [6].

Homozygous Df 2R Dg248 clones were generated by using ey-FLP/FRT [30] system.

T cell clones were generated by limiting dilution cloning in 96-well round-bottom plates.

Clones were generated using the Flp recombinase system described previously [52].