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Quadratic equation was obtained for freeze-dried samples applied by 3 cycles of microwave with a regression coefficient of 0.83.
Linear equations were obtained for oven-dried samples applied by 1 and 3 cycles of microwave with regression coefficients of 0.65 and 0.62, respectively.
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Samples were applied by CAMAG Linomat 5, auto-sampler (Switzerland) using CAMAG micro-syringe, 100 µL (Switzerland) on HPTLC aluminum plates, pre-coated with silica gel 60 F254 (20 × 20 cm) (Merck, Germany), 200 µm thickness and 5 µm particle size.
The samples were applied by an autosampler (Perkin Elmer).
The protein samples were applied by rehydration loading into the IPG strip.
The column was pre-equilibrated in 10 mM TEAB buffer, and samples were applied by syringe at a flow rate not exceeding 30 mL min 1.
Random sampling was applied by using Stata 11 (StataCorp LP, College Station, TX, USA) with a representation of samples for each year.
Concentrations of TNFα were estimated by the reduction of the optical density of control wells by unknown samples applying a standard curve generated by standard concentrations.
Future sampling with ichthyocides applied by divers utilizing rebreathers, supplemented with trawl and dredge sampling would allow collecting from the outer slopes and would certainly yield additional new and interesting species of fishes from this submerged atoll.
A probability-based sampling approach was applied by Hansen et al. [4] that employs MODIS data to identify areas of likely forest cover loss and to stratify probability of forest clearing.
Furthermore a third procedure for crude sample fractionation was applied by sequential NH4SO4 precipitation (40 %, 70, and 90% NH4SO4).
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