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According Table 6, the two best models are farnesyl diphosphate synthase and dihydroorotate dehydrogenase, with tenfold cross validated specificity values of 0.980 ± 0.016 and 0.870 ± 0.042, respectively.
In addition, we have validated specificity and sensitivity of this robust reporter system.
We validated specificity of the antibodies by immunofluorescence in the undifferentiated human embryonic stem cell (hESC) lines, H9 (XX) and HSF1 (XY) (Fig. 3A).
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At the end of each run melting point analysis was performed to validate specificity of PCR transcripts.
When SYBR analysis was performed, cycling reactions were followed by a dissociation curve analysis to validate specificity of amplified products.
To validate specificity of the anti-PTEN antibody, 3-dpf-old ptena −/−and ptenb −/− (Faucherre et al., 2008) and control embryos were lysed in cell lysis buffer.
Samples of PCR using primers directed against p21 cyclin, cyclin G1, socs3a and gapdh were analyzed by agarose gel electrophoresis to validate specificity of amplification products.
Isotype-matched negative control antibodies and negative control mouse IgG2a, kappa (DakoCytomation) were used to validate specificity of Ki-67, p27KIP1, and EGFR staining.
Note that, to validate specificity of the probes targeting GGGGCC and CCCCGG RNA, FISH was carried out on frontal cortex and cerebellar tissue from ALS, frontotemporal lobar degeneration (FTLD), and FTLD-MND patients with normal C9ORF72 repeat length.
Dot blotting analysis was performed to validate specificities and sensitivities of the respective monoclonal antibodies.
The absence of staining in Ehd3 /– glomeruli validated the specificity of GEnC-specific EHD3 expression (Figure 1B, D and F).
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