The mean leukocyte TL was assessed using the TRFL method, which was measured using the Southern blot method in duplicates.
In order to assess the utility of the above protocol, we performed a pilot screen of 7368 compounds (derived from a variety of different compound libraries, see Methods) in duplicates for a total of 44 384-well plates.
We calculated the mRNA copy number with a standard curve method, in which duplicates of recombinant plasmids with known copy number representing each globin cDNA were run using serial dilutions 10-fold (10 10).
Hemoglobin concentration (g dL−1) and hematocrit, using the micro-hematocrit method, were measured in duplicate and then used to estimate percentage changes in PV [6].
The method was conducted in duplicate for each organism evaluated.
Each method was performed in duplicate, using two different peppers from one batch.
Total RNA was extracted by TRI-reagent/chlorophorm method, then assayed in duplicate using a Rotorgene 3.0 Real Time PCR instrument (Corbett Research, Cambridge, UK) and the Real Time PCR amplification kit SYBR Green I (Qiagen, Crawley, UK).
Additionally, some minor changes to the method were tested in duplicate: (1) extraction for 30 min instead of 15, (2) dry the PBA cartridges by applying vacuum for 1 min after applying the extract and after washing the cartridge and (3) evaporation of the solvent at 55 °C instead of 50 °C.
All measurements with the three methods were performed in duplicate.
Pairs of reviewers with training in research methods, independently and in duplicate, screened study reports for eligibility, assessed risk of bias, and collected data from each eligible study.
