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The only other manual steps required for extraction set up were the addition of elution buffer, collection tubes and plungers into the cartridge-holder.
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Blood samples were taken into EDTA buffered blood collection tubes (Sarstedt, Germany) and processed with standard procedures by Ficoll-Paque (GE Healthcare, USA) dense-gradient centrifugation to isolate buffy coat.
The sampled peritoneal lavage fluid was centrifuged at 400×g for 5 min and then washed with the collection buffer.
Study Design: Genomic deoxyribonucleic acid and total ribonucleic acid were extracted from cell pellets obtained from the residual fluid-based Papanicolaou specimen collection buffer after clinical processing.
Cells were washed twice in ice cold PBS and suspended in collection buffer (100 mM Tris-HCl pH 9.4, DTT 10 mM Protease inhibitors).
All subjects were required to rinse their mouths with saline buffer before sample collection.
Cells were washed two times with PBS and resuspended in 1 ml of collection buffer.
A surgical laparotomy was performed, and the uterine horn was removed and placed into ice-cold tissue collection buffer.
The time elapsed until actual usage of the buffer for sample collection ranged from a few weeks to two months and we observed a decrease in volume in some of these tubes, probably due to evaporation.
Tissue samples were placed in lysis buffer immediately after collection, and total RNA was extracted by PureLink RNA Mini Kit (Ambion, USA) following the procedure specified by the manufacturer.
A sample of glass eels for each site was fixed for 24 h at 4°C in a saturated salt buffer immediately after collection in the field (T0) to preserve RNA and DNA integrity.
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