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For a sufficient amount of sediment material each sample from the open cores (SAT 7 and 10) represents sections of 5-10 cm from significant stratigraphic units.
For these tissues, due to the small amount of material, each sample represented a pool of three young plants.
Single-stranded, then double-stranded cDNA was synthesized from the poly(A)+ mRNA present in the isolated total RNA (10 μg total RNA starting material each sample reaction) using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen Corp., Carlsbad, CA) and poly (T -nucleotide primers T -nucleotided a sequence recognized by T7 RNA primersase.
In this survey the considerable quantity of mixed helminthic species often present in the same intestinal content could have affected the equal distribution of the material (each sample was divided in aliquots for all the examinations), hampering thus the performance of the first PCR in the detection of M. lineatus and T. taeniaeformis DNA.
Double-stranded cDNA was synthesized from the poly(A) + RNA present in the isolated total RNA (8.5 g total RNA starting material each sample reaction) using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Carlsbad, CA, USA) and poly (T -nucleotide primers T -nucleotided a sequence recognized by T7 RNA primersase.
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The team measured the amount of organic material in each sample and isolated and identified amino acids--the building blocks of proteins--as well as amines--the products of amino acid degradation.
All the lyophilized material for each sample was dissolved in 0.65 mL each of D2O and H2O, 1.35 mL of the NaOH-EDTA extracting solution and 0.8 mL of 10 M NaOH.
For electrical conductivity measurements and sensing experiments, 0.2 g material from each sample was palletized at room temperature with the help of a hydraulic pressure instrument at 25 KN pressure for 10 min.
This allowed us to obtain accurate measurements of the surface concentration of metals using only 1 ({mathrm{cm}^2}) of material from each sample, which is a relatively small amount compared to the total area available (each paper sample measured approximately 35 ({mathrm{cm}^2})).
Fresh weight was determined and 2 g of fresh material of each sample were dried at 60 °C for approximately 72 h, until constant weights were achieved; tissues were weighed to estimate the factor for conversion from fresh to dry weight of each replicate.
Extractions were performed with 0.1 g dry material for each sample with three replications.
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material each test
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