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To determine the usefulness of the Pentacam corneal volume assay in the assessment of corneal endothelial damage caused by phacoemulsification and aspiration (PEA).
cDNA was analysed in a 25 μl final volume assay system containing 300 nmol l−1 primers and 200 nmol l−1 TaqMan hybridisation probe (Biosource UK Ltd).
The optimal primer concentration of each forward and reverse locus-specific primer was determined in preliminary PCR assays varying the primer concentration between 5 and 120 nM (Table 1) and also was included within each 10 μL (total volume) assay.
Sugar content were determined by the anthrone method described by Spiro [ 70]. 100 μl leaf extract were added to 3 ml (final volume) assay media containing 1.08 M H2SO4, 1.09 mM thiourea and 2.1 mM anthrone.
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Such valves constructed from non-adsorptive poly-PEGDA could also find use as pumps, for application in small volume assays interfaced with biosensors or impedance detection, for example.
With the exception of the 200 µL volume, assays for Fdh [26], MtdA [42], Acn [43], were as described previously.
In order to account for these small differences, the amount of bone formed was normalised to the volume assayed (BV/TV).
The beauty of microfabrication is that such tiny volume assays can be repeated many times, in parallel, on a single microchip.
Twenty five μL final volume assays contained: 5 μL of cDNA, 0.5 μL of primers (200 nM each), 0.25 μL of probes (200 nM each), 12.5 μL of QuantiTect Multiplex PCR Mastermix (Qiagen, Courtaboeuf, France) and 3.75 μL of water.
More specifically, we used the patch clamp to sample from the cytoplasm and perform a small-volume assay using CE MS.
Effective studies of clinical specimens require low-volume assays, high precision measurements, and the ability to process many samples.
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