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Therefore, to separately count each strain the total number of cells were counted, samples were then incubated, for 5 min in 0.5% Triton X-100 (Sigma, USA), to completely lyse the cell wall deficient mutant cells.
Cell viability in counted samples was determined by annexinV binding (Invitrogen, Carlsbad, CA, USA) and by propidium iodide exclusion.
Hence, the experimental procedure started with isolation of total RNA from 9 independent and cell counted samples drawn from the same cell culture.
We only counted samples as "cleaved" if the unheated control sample showed significant PstI digestion or, in a few cases, if the unheated sample did not PCR amplify, then a sample was counted as "cleaved" only if greater than 80% were not digested by PstI.
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When liquid samples are involved, this effect can usually be minimized by consistently counting samples of equal volume.
Methods for counting samples under conditions of secular equilibrium and non-equilibrium between 32Si and its daughter isotope 32P are described.
Identifications when counting samples were only made to the level of functional group, as described above, and not to species level.
Too numerous to count samples were assigned 1.5 times the smaller volume upper detection limit.
Radioactivity was determined by counting samples using a Beckman LS5801 counter (Beckman Instruments, High Wycombe, Buckinghamshire, UK).
MFSelector does not count samples on the wrong side of the discriminating line more than once.
To do this, we overlaid the telemetry data on hypothetical project footprints and simulated a variety of point count sampling strategies for those footprints.
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