Total RNA from myocardial tissues (100 ng per specimen) was combined with 200 nM each of forward and reverse primers, 1x One-Step RT-PCR master mix and, 100 nM of TaqMan probe in a 50 µl reaction.
For unextracted samples, 1 μl of clinical specimen was combined with 4 μl of deionized water, 0.5 μl of internal control DNA and 15 μl of PCR Master Mix.
HC2 testing was conducted per the manufacturer's instruction, with one exception: 50 μl of the DCM specimen was combined with 25 μl of the kit denaturation reagent rather than combining 1000 μl of STM specimen with 500 μl of the kit denaturation reagent.
Variation of copper concentrations was generally higher in the experiment at ECN than at BAM - perhaps due to the fact that a single test specimen per test assembly was used in the experiments at ECN whereas two test specimens were combined in the test assemblies at BAM (see Table 3).
Aliquots (1 ml) of all prospective and retrospective specimens were combined with A549 cells (Viromed Laboratories, Minnetonka, MN) in shell vials and placed at 37°C.
These specimens were combined in a tube of veal infusion broth transport medium and delivered at ambient temperature within 1 to 2 h to the site research laboratory.
The partial mtDNA cytb gene and the D-loop sequences from ancient Chinese specimens were combined with previously published sequences for network and phylogenetic analysis.
These specimens were combined with previously-collected samples from Lakes Victoria, Albert and Kivu [ 6] stored in Axel Meyer's collection at the University of Konstanz.
Studies were excluded if they were not in the English language, if they referred only to respiratory specimens, or if the results of respiratory and non-respiratory specimens were combined and could not be disaggregated.
In the two cases where patients were stratified by immunophenotype (Pieters et al, 1998; Kaspers et al, 2005), data from T- and B-lineage specimens were combined before analysis.
For each species, the sequencing reads from the three specimens were combined into a single data set and de novo assembled using Trinity (Grabherr et al. 2011) with default settings to generate the "reference assembly".
