Genome Engineering by Transgene- Instructed Gene Conversion in C. elegans - Caenorhabditis Elegans: Molecular Genetics and Development - page 73

Fig. 3 (A) Description of a standard MosTIC repair template. (B) Repair templates and control
plasmids for the engineering and PCR screening of MosTIC KO/del and KI alleles. Primers required
for construction of the templates and PCR screening are described in the text.
polymerase. If necessary, a gradient of annealing temperature ranging from 55 to
68 C can be performed to optimize the amplification of individual and fused frag-
ments. After amplification, PCR products are purified by gel extraction using the
Qiaquick Gel Extraction Kit (Qiagen) according to the instructions of the manufac-
turer. The final PCR product corresponding to the full-length repair template is
subcloned using the Zero Blunt PCR cloning kit (Invitrogen) or any other cloning kit
and sequenced.