Moreover, both ASCs (isolated due to MAL and UAL) were characterized by high expression of markers specific for ASCs: CD9, CD10, CD34, CD44, CD49d, CD54, CD55, CD59, CD71 and low expression of CD11a, CD11c and CD144 which are described as a non-specific ones (Table 3).
SNP genotyping was performed by comparison of Ct values with a homozygous sample having a lower Ct with the specific primer set and a higher Ct with the non-specific one, and a heterozygous sample having approximately equal results from both assays.
In such cases, if there is no other tissue sample, it is necessary to confirm the specificity of the DNA product either by hybridization analysis or by using a nested primer set that will only amplify a specific DNA product and not the non-specific one.
Specific hybridization competes with non-specific one.
Hence, specific hybridization starts to dominate over non-specific one always at the beginning of the mix-range.
This S/N-ratio, R, provides the specific binding strength of the PM in units of the non-specific one.
In this analysis we found that reducing the fixation time to 30 min at RT for 24-hpf embryos significantly enhanced the specific signal, while reducing the non-specific one (Additional file 5H).
Although most proteins in the eluates appeared to be non-specific, one protein band was remarkably enriched in eluates associated with H2A.Z-H2B dimers but not in eluates associated with H2A-H2B dimers.
It thus defines a characteristic S/N-ratio (in logarithmic scale) of the chip which characterizes log-ratio of the mean specific and non-specific signals, or in other words, to which extend the specific signal exceeds the non-specific one.
In our experimental system, by using the K-17 antibody, we discriminated the specific DAX-1 band of ∼50 kDa MW, from the non-specific one at 60 kDa, as they did in Helguero et al. As shown in Supplementary Figure 1, specificity of the ∼50-kDa band was proved by DAX-1 silencing experiments.
