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A virus titration in the absence of horse serum was made to confirm that the virus dose per well was 100 TCID50.
A virus titration was included to check the virus dose of the assay was correct and a titration of horse antisera positive for AHSV-4 VN antibodies were included in the assay as positive controls.
In the samples derived from all 5 passages, the virus titer (CCID50/mL) was determined by a virus titration assay.
Although 0.05 mM ribavirin failed to mediate significant suppression of VR2332 replication, concentrations of 0.1 and 0.2 mM ribavirin were able to suppress virus replication to undetectable levels based on a virus titration assay at passage 2. However, virus replication started to resume at passages 5 and 17 in the presence of 0.1 and 0.2 mM ribavirin, respectively.
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Virus titration analysis showed a clear correlation between GFP signal and the number of infectious virus particles produced in different cell lines under various treatment conditions (Fig. 1C).
The mean plaque area of each virus yielded a characteristic size distribution, regardless of virus titration (data not shown).
Virus titers in serum were determined by virus titration on PAM following a standard procedure [ 32]. 24-h cultivated PAM were inoculated with 10-fold dilution series of the serum samples.
For each supernatant sample, cells or debris were removed by a low speed centrifugation and the supernatant was used for virus titration by TCID50 on Sf9 cells.
The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration.
Three days post-virus infection, virus titration, real time RT-PCR, and RT-PCR assays were performed to verify the effect of siRNA in AMPV replication.
The half-life of ASFV in the air was on average 19 min when analysed by PCR, and on average 14 min when analysed by virus titration.
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