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198 Caldwell et al.Fig. 1. (A) Protein X is made up of two exons. Therefore, the deletion of exons Aand B is all that is required to create a knockout. (B) Protein X is made up of threeexons. Exon C does not contribute significantly to the protein and is made up of almostcompletely of the 3′UTR. Therefore, the deletion of exons A and B is all that is requiredto create a knockout.tried to reverse the sizes of the upstream and downstream arms. If this does notsolve the problem, the arms might be shortened down to 1 kb. If this adjustmentfails, the positions of the arms within the locus need to be shifted. Other problemsare PCR amplification failures or difficulties to clone an arm sequence. Ifthe failure involves an upstream arm, pairs of new upstream primers aredesigned 500 bp closer to the downstream primers as long as the arm sequencestill exceeds 1 kb. In addition, a pair of new downstream primers is made at thenext suitable position. If the failure involves a downstream arm, pairs of newdownstream primers are stepwise designed 500 bp closer to the upstream

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