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To reduce risk of nosocomial transmission samples were collected in isolation in a ventilated room with the supervising physician using a personal respirator.
Cell suspension was filtered through a sterile nylon mesh and collected in isolation medium [DMEM, 10% FBS, 50 mg/ml gentamicin, 2 mM l-glutamine, 20 ng/ml epidermal growth factor (EGF)].
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Cells were collected in mitochondrial isolation buffer (250 mM Sucrose, 10 mM Tris-HCl pH 7.4, 0.1 mM ethylene glycol tetraacetic acid (EGTA)) and homongenized on ice using a Dounce homogenizer.
Furthermore, more conidia were collected in the new isolation unit than using a standard method.
Also, because the tissues were collected in RNAlater, virus isolation attempts could not be performed.
With the patient's consent, saliva and throat wash (by gargling 10 mL normal saline) were collected in an airborne isolation room, according to the guidelines for aerosol-generating procedures (17 ).
Blood samples were obtained by venipuncture and collected in EDTA vacutainers for plasma isolation and in coagulation tubes for isolation of serum.
Total genomic DNA was extracted from tail fin or tail muscle from male fish collected in 2001 using standard phenol/chloroform isolation [ 36] or using the PURGENE DNA Isolation Kit (Gentra Systems, Inc., Minneapolis, MN, USA).
Using a scalpel blade, the tumor foci from the marked areas were scraped and collected in micro tubes for DNA isolation.
Peripheral blood was collected in EDTA for genomic DNA isolation from leukocytes of all affected and unaffected members.
Tissues for RNA isolation were collected in a commercial sheep slaughterhouse, immediately frozen in liquid nitrogen, crushed to powder the same day and stored at -80°C.
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