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Viral <t>nucleotide</t> loads in growth curve cultures. (A) The dynamic range and linearity of the CVA12 real-time assay. The threshold cycle (Ct) values (x-axis) were plotted against tenfold dilutions of CVA12. The parameters of the slope and y-intercept are given by the equation. R 2 , regression coefficient. (B) The levels of CVA12 RNA in the culture supernatants at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (C) The levels of CVA12 RNA in the cell lysates at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (D) Virus titer detection in RD and Hep-2 cell lines infected with 100 TCID 50 of CVA12 stock. The culture were collected at 6, 12, 24, 48 and 72 h post-infection (hpi). All the data are expressed as the mean ± SD of three biological replicates. Statistical significance was determined by ordinary one-way analysis of variance (ANOVA) with multiple comparisons methods using the GraphPad Prism software. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
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Viral nucleotide loads in growth curve cultures. (A) The dynamic range and linearity of the CVA12 real-time assay. The threshold cycle (Ct) values (x-axis) were plotted against tenfold dilutions of CVA12. The parameters of the slope and y-intercept are given by the equation. R 2 , regression coefficient. (B) The levels of CVA12 RNA in the culture supernatants at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (C) The levels of CVA12 RNA in the cell lysates at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (D) Virus titer detection in RD and Hep-2 cell lines infected with 100 TCID 50 of CVA12 stock. The culture were collected at 6, 12, 24, 48 and 72 h post-infection (hpi). All the data are expressed as the mean ± SD of three biological replicates. Statistical significance was determined by ordinary one-way analysis of variance (ANOVA) with multiple comparisons methods using the GraphPad Prism software. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Frontiers in Microbiology

Article Title: The genetic and proliferation characterization analysis of novel coxsackievirus A12 in Beijing, China

doi: 10.3389/fmicb.2025.1665461

Figure Lengend Snippet: Viral nucleotide loads in growth curve cultures. (A) The dynamic range and linearity of the CVA12 real-time assay. The threshold cycle (Ct) values (x-axis) were plotted against tenfold dilutions of CVA12. The parameters of the slope and y-intercept are given by the equation. R 2 , regression coefficient. (B) The levels of CVA12 RNA in the culture supernatants at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (C) The levels of CVA12 RNA in the cell lysates at 6, 12, 24, 48 and 72 hpi were determined by qRT-PCR ( n = 3 biological replicates). The 100 TCID 50 of CVA12 stock were used to infect RD and Hep-2 cell lines, respectively. (D) Virus titer detection in RD and Hep-2 cell lines infected with 100 TCID 50 of CVA12 stock. The culture were collected at 6, 12, 24, 48 and 72 h post-infection (hpi). All the data are expressed as the mean ± SD of three biological replicates. Statistical significance was determined by ordinary one-way analysis of variance (ANOVA) with multiple comparisons methods using the GraphPad Prism software. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: A nucleotide identity matrix was generated using BioEdit software.

Techniques: Quantitative RT-PCR, Virus, Infection, Software

The phylogenetic characteristics of CVA12. (A) Nucleotide distances between the different groups and within each group. (B) Principal components of CVA12 sequences with genogroups used as prior groups based on VP1 coding region. Different colors represent prior genogroups, and individual sequences are marked as dots. Results of eigenvalues analysis (PCA and DA) displayed in the inset. Axes represent the frst two principles. PCA, principal components analysis; DA, discriminant analysis. (C) Maximum likelihood phylogenetic tree based on the full-length VP1 coding region, with 1,000 bootstraps and SH-like approximate likelihood ratio tests (SH-aLRT) replicates at each node. The scale bar indicates nucleotide substitutions per site per year. The red colors represent the five CVA12 strains in this study.

Journal: Frontiers in Microbiology

Article Title: The genetic and proliferation characterization analysis of novel coxsackievirus A12 in Beijing, China

doi: 10.3389/fmicb.2025.1665461

Figure Lengend Snippet: The phylogenetic characteristics of CVA12. (A) Nucleotide distances between the different groups and within each group. (B) Principal components of CVA12 sequences with genogroups used as prior groups based on VP1 coding region. Different colors represent prior genogroups, and individual sequences are marked as dots. Results of eigenvalues analysis (PCA and DA) displayed in the inset. Axes represent the frst two principles. PCA, principal components analysis; DA, discriminant analysis. (C) Maximum likelihood phylogenetic tree based on the full-length VP1 coding region, with 1,000 bootstraps and SH-like approximate likelihood ratio tests (SH-aLRT) replicates at each node. The scale bar indicates nucleotide substitutions per site per year. The red colors represent the five CVA12 strains in this study.

Article Snippet: A nucleotide identity matrix was generated using BioEdit software.

Techniques: