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The procedure was designed so that all three analyses could be performed starting with a single sample given the difficulty of preparing multiple aliquots in known ratios.
Coded samples are received by the facility and processed into appropriate fractions (e.g., plasma, mononuclear cells, granulocytes, total white blood cells and red blood cells) and frozen in multiple aliquots in more than one freezer whenever possible.
Blood samples were centrifuged, IEs were buffy coat depleted, washed three times in phosphate-buffered saline (PBS) and cryopreserved in multiple aliquots in glycerolyte solution at −80°C.
The isolate was stored as multiple aliquots in skim milk (Oxoid Ltd, London, UK) under -70°C.
The separated plasma samples were stored in multiple aliquots in a freezer at −80°C until assays.
The isolate was stored as multiple aliquots in skim milk (Oxoid Ltd, London, UK) at -70°C.
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Purified RNA samples from both the blood and dura were stored in multiple aliquots at -80°C in order to minimize the number of freeze-thaw cycles.
Collected serum or plasma was stored in multiple aliquots at −75°C until assayed (in duplicate).
DNA from all samples is archived in multiple aliquots at -80C at CSRIO Oceans and Atmosphere in Hobart, Tasmania and is accessible for re-analysis should the proposed methods be deemed to provide a substantial improvement or progress over prior results.
Cells were frozen down in multiple aliquots at passage 3 to 7, and stored in liquid nitrogen until use.
l-[1-C]Ascorbic acid was dissolved in 0.1 mM acetic acid and stored in multiple aliquots at −20 °C.
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