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While using the same cycles of hydrogenation and dehydrogenation, heat shall be stored on the contrary by high reaction enthalpy in the LORC.
PCR were performed in a Robocycler Gradient 96 apparatus (Stratagene) using the same cycles as for round #1.
This was accomplished by using 10ng of the VL and VH PCR products along with the 'scforward' and 'scback' primers (see below) in a PCR reaction using the same cycles and reaction conditions as before.
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The second round PCR (25 cycles) was done using the same cycle schedule.
Cycling was carried out using a Biometra trio-thermblock (Biometra®) with 35 amplification cycles consisting of denaturation at 94°C for 30 sec, annealing at 60°C for 30 sec and extension at 72°C for 1 min. For the second round of this nested PCR, 3 μl from the first PCR reaction were amplified with the inner pair of primers using the same cycle as before.
This means using the same cycle that you would use to wash wool.
Three microliters of RT-PCR reaction were added to the second step (nested) PCR and amplified for other 30 cycles, using the same cycling conditions of the first step.
The fluorescent labelled primer was added in the secondary reaction using the same reaction cycles.
All exercise tests were made using the same bedside cycle ergometer (Fig. 1) (MOTOmed Letto 2, RECK-Technik GmbH and Co. Betzenweiler, Germany).
We have also examined healing for shorter periods, as little as 30 min, using the same temperature pressure cycle, arriving at essentially the same final results, i.e., full toughness recovery.
These results indicate that the Geobacter species and their last common ancestor all oxidized acetate using the same TCA cycle pathway that produces NADH, NADPH, and reduced ferredoxin.
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