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However, the two assay sets have very similar averages and standard deviations to each other, and hence the analysis and discussion below, which are based on the 1,528 assay set, also hold for the 843 assay set.
The results from the 843 assay set are included in Additional file 4. The similarity score distributions from the two assay sets have very similar averages and standard deviations to each other, and hence, the conclusions drawn from the 1,528 assay set should also be valid for the 843 assay set.
Therefore, a subset of the 1,528 assays were generated by selecting 843 assays which have active compounds only (without any inconclusive or unspecified compounds), and the same analyses were performed both for the 1,528 assay set and the 843 assay set.
Three endogenous controls were added to the assay set.
On the day of assay set up, cell count and viability were determined using a Guava Cell Counter.
A comparable inhibition occurred in the HALO-Hemogenix luminescence assay, set up with different growth factor cocktails to reveal proliferation of progenitors with distinct differentiation potentials (CFU-C, CFU-C+BFU-E and CFU-C+BFU-E+CFU-GEMM) (Fig. 2C).
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Commercially available reagent mixes facilitate PCR assay set-up with fewer steps and timeliness.
The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses.
Further, the role of solubility as a governing factor for colloidal aggregation formation within a specified assay setting is examined and considered as a possible cause of a high cross-assay activity score.
Assay set-up is diagramed in Figure 2A.
The SYBR Green methodology seems to be a better option for quantification than compared to sequence-specific fluorescent probes in the TaqMan or TaqMan MGB based assay, since it requires only one set of specific primers, hence providing additional experimental flexibility, and it reduces assay set-up and running costs while providing similar levels of accuracy in optimised assays [25].
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