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Constitutive negative/positive, point/sequential mutations involving residues Thr-210, and Lys-238 in WT TR2 (CMvector/Gas4) vectemplateemplate were made according to QuikChange XL site-directed mutagenesis kit (Stratagene) as described previously [PNAS].
Changes to the template were made through an iterative process before implementation.
Several modifications to the template were made during the study period.
The plasmid DNAs containing a single target or competitor template were made by TA cloning of the corresponding PCR amplicons into pCR2.1 (Invitrogen) so that the inserted fragments could be liberated with EcoRI restriction.
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The template was made in the United States, a counternarcotics strategy originally designed for Colombia.
In contrast, when a repair template is made available during the attempted gene repair process, the new information that the repair template carries becomes incorporated into the DNA such that the target gene is either repaired (if it was defective) or acquires a new function.
A template was made to allow for quick hole placement.
Decision on the vertical movement of deformable template is made by sine functions (Figure 10).
A template was made with a hole-punch matched to the diameter of the spectrophotometer.
Decision on the horizontal movement of deformable template is made by cosine functions (Figure 8).
Accordingly, a deformable parametric template is made evolve by internal and external forces.
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