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Using a single technician for this process with space, equipment and reagents that are not used for other purposes is also critical.
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Using a single booster.
Using this process a single technician can amplify, pool and adapt up to 96 samples in two days.
Using these methods a single technician can process up to 96 samples at once, creating a sequence-ready library in under three days.
Despite these limitations, strengths of the current study include the inclusion of unaffected participants with a known BRCA1 mutation status and the use of a single technician (who was blinded to the intervention group) who analysed all of the samples from the same participant in a single batch under the same conditions.
All samples were analyzed by a single technician using the same equipment throughout the study period in a reference laboratory.
Three independent assay runs were performed for each sera pool per virus by a single technician using the method detailed below: In brief, serial 10-fold dilutions of the serum pools were mixed with a constant challenge dose of each respective JEV (45 60 plaques per well) and inoculated in duplicate, onto wells of a 6-well plate of confluent LLC-MK2 cells.
A single technician assessed sperm morphology using the strict morphology method, as recommended by the WHO (1999).
Sperm morphology was assessed from smears after Shorr staining, using the modified David classification [ 17], by a single technician at the Reproductive Biology Laboratory, Cochin University Hospital, Paris.
A single technician scored sperm concentration in duplicate using disposable counting chambers (Cellvision, Heerhugowaard, the Netherlands), and determined sperm morphology on air-dried Papanicolaou-stained seminal smears.
Arterial stiffness was measured in the left common carotid artery using a portable Acuson Cypress (Siemens Medical Solutions USA Inc, Mountainview, California, USA) platform with a 7 MHz linear-array transducer by a single technician.
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