Thus, the assays worked well even when only 1 ng of DNA was used.
Pilot studies demonstrated that the assays worked with as little as 1 ng of DNA (Fig. 3).
The trypan blue and LDH assays work on the same principal, porous membranes allowing the passing of molecules.
Consequently, in our study, the initial subtraction assays worked as small-scale screenings for potential transcripts.
Principally the assay work by converting lactate to pyruvate through oxidation by the LDH, subsequently Pyruvate reacts with the tetrazolium salt to form formazan, water-soluble formazan dye that can be detected by a spectrophotometer [21].
The assay worked accurately with 100% specificity in at least three real time PCR platforms.
The assay works on the principle that when Annexin-V bounds to phosphatidylserine, it shows increased fluorescence and hence indicates the externalization of the plasma membrane.
As often happens when creative people work together in this way, they had a lucky break within a couple of weeks and had the assay working in manufacturing a short while later.
Surprisingly, neither the murine TAAR3 nor the other TAAR3 orthologs tested displayed cAMP production following incubation with cyclohexylamine (Figure 5A) and isoamylamine (data not shown) although the assay worked properly with TAAR4 and TAAR5 (Figures 5B, C).
