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Relative virus accumulation was calculated using efficiency adjusted ΔΔCt methodology, incorporating the reference transcript to control for variation in loading [37], [38].
Ct values are a semiquantitative measure of virus load and therefore only reflect relative virus loads.
IPNV gene used for relative virus quantification (described in Results section).
Transduction index (TI = % GFP+ cells x geometric mean fluorescence intensity) is a linear indicator of relative virus activity.
However, to confirm the relative virus titres, the virus concentration was also assessed by QPCR (Supplementary Figure S2 at http://www.bioscirep.org/bsr/033/bsr033e003add.htm).htm
The relative virus detection rates from different specimens during the illness suggests that respiratory specimens (nasopharyngeal aspirate, throat swab) are more useful in the first 4 days of the illness, while fecal samples are better later in the illness.
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This entry defect may have contributed to our observation of lower vaginal and spinal cord titers at some time points after infection with viruses containing gD mutations relative to virus with wild-type gD (Fig. 4).
Since pretreatment and co-treatment of mice with PT (relative to virus inoculation) increased lung viral loads, but not PT treatment after virus inoculation, this indicated that PT is targeting a component of the early innate immune response with an important activity within the first 24 h of virus infection.
Immobilized lentivirus produced large numbers of transduced cells, and increased transgene expression relative to virus alone.
Serum antibodies with the capacity to neutralize LCMV appear with delayed kinetics relative to virus specific IgG in wild-type mice [ 26].
Titers were reported as the reciprocal of the greatest serum dilution resulting in a 90% or greater reduction in FFU relative to virus control well.
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