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Despite using a different method, Kit A showed qualitative results (animal classified as positive or negative) very similar to the other two kits.
For the determination of l- and d-lactate concentrations in the co-culture media, we used the d-lactic acid/l-lactic acid (UV method) kit of R-biofarm (Roche, Vilvoorde, Belgium) as described previously (De Ryck et al. 2014).
Chromatin immunoprecipitation (ChIP) was carried out using enzymatic shearing method kit (Active Motif, Carlsbad, CA, USA) according to the manufacturer's recommendations with the following antibodies: mouse anti-CREB (Invitrogen, Carlsbad, CA), rabbit polyclonal anti-ATF2 (Santa-Cruz, Santa Cruz, CA, USA), rabbit polyclonal anti-MeCP2 (Upstate, Chicago, IL, USA).
Total intracellular TG content was measured using a fluorometric method kit (BioVision, CA, USA) in accordance with the manufacturer's instructions.
Blood was obtained by intracardiac puncture, serum was separated by centrifugation, and the amount of total protein was measured using the bicinchoninic acid method kit (Pierce).
Histochemical staining for collagen fibers in uterine tissues was performed using the ACCUSTAIN Trichrome Stains (Gomory method) kit (Sigma) according to the manufacturer's instructions.
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Starting with 10 µg of total cellular RNA, Eberwine method (kits from Roche) was used to generate cDNA and thereafter Cy3 and Cy5 (Amersham) labeled antisense RNA.
Glucose was determined in serum by colorimetric assay method kits (Biodiagnostic Chemical Company, Cairo, Egypt).
In our experience, of the five one-step method kits tested, we found three that yielded satisfactory analytical sensitivity with an LOD 95% of 15 50 copies/ µL.
Starting with 10 μg of total cellular RNA, Eberwine method (kits from Roche) was used to generate cDNA and thereafter Cy3 and Cy5 (Amersham) labeled antisense RNA.
The genotyping performance using wgaDNA made by the two OmniPlex method kits, GPlex2 and GPlex4, was independent of the DNA-extraction method, with GPlex4 showing consistently higher call rates than GPlex2 (Wilcoxon's paired test, p < 0.001).
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