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For each primer pair, melting curves were analyzed to validate specificity of binding.
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.
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Dot blotting analysis was performed to validate specificities and sensitivities of the respective monoclonal antibodies.
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).
To further validate the specificity, 100 positive cytotoxin-associated protein (CagA) samples were also collected to test the specificity of the reader.
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