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The functionality of the fingers is compromised because of the bone alterations and the thickening of the subcutaneous tissues, resulting in a failure of a complete extension of the fingers with a "claw hand" deformity [19, 20].
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It has turned out subsequently that the axiom V = L gives a virtually complete extension of ZFC.
The criterion for a positive PER was complete extension of the proboscis.
The amplification conditions were as follows: 2 min of initial denaturation at 95°C, then 40 cycles at 95°C for 30 s, 50°C for 30 s, and 72°C for 1 min. The amplification was completed by a final step of 5 min at 72°C to allow complete extension of all PCR products.
A final 7-min 72°C step was added to allow complete extension of the products.
The following temperatures were used for cycling: 94°C for 4 min followed by 30 cycles at 94°C for 30 s, 50°C for 30 s and 72°C for 1 min 30 s followed by a final step at 72°C for 40 min, to ensure complete extension of all products to maximize TA cloning.
A final incubation of 72°C for 4 min was added to ensure complete extension of amplified products.
The PCR cycling conditions were 5 min at 94°C followed by 35 cycles of 45 s at 94°C, 45 s at 66°C and 45 s at 72°C and with a final step at 72°C for 15 min to allow for the complete extension of all PCR fragments.
6) Extend the signal at the right endpoint in the same way which is shown in step 5, and get the complete extension of the signal as (x'(t)).
When this distraction was accomplished, the tibial clamp was locked and distraction now applied to the posterior unit (at 1 mm/8 h), with the femoral clamp on the rail and hinge unlocked, until complete extension of the knee was achieved.
A final 5-minutes elongation at 72°C was performed after the last cycle to ensure complete extension of the amplicons.
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