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The fragment was purified and sequenced as indicated below for all other sequenced PCR products (GenBank accession: Feeding trial: EF590264, EF590265, Sarasota fish samples: EU239803 EU239814, EU239933).
All RT-PCR, except as indicated below for amplification of the NAIP ORF and generation of the expression vector, was performed with Platinum Taq DNA Polymerase (Invitrogen) and the relevant primers are listed in Table S1, all used at 10 µM.
Using 5 Prime HotMasterMix (Eppendorf), PCR reactions were performed as follows: 94°C for 2 min, then 32 cycles 30 sec denaturation at 94°C, 30 sec of annealing (as indicated below for each primer pair), and 30 sec of elongation at 65°C, then a 2 min final elongation at 65°C.
The vessels were allowed to equilibrate for 1 h before being pretreated with pharmacological agents indicated below for 20 min.
B. cenocepacia was subcultured to obtain an initial optical density at 600 nm (OD600) of 0.02 and grown for the time indicated below for each experiment without selection.
For single antibody staining, individual sections were incubated with primary antibodies (diluted as indicated below) for 40 min at room temperature (anti-SV40 and anti-securin antibodies were incubated overnight at 4°C).
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Immunomonitoring studies were performed for the number of individuals indicated below, that is, for the subjects whose frozen blood samples were properly stored and had a cell viability superior to 90% on thawing.
The Chr X haplotype for each male in the vicinity of the Gct4 locus is represented as a vertical column, while the overall GC tumor incidence observed in the F1 daughter offspring is indicated below the haplotype for each tested male.
The objective was to test hypotheses of different ω values (ω is the ratio of nonsynonymous substitutions per nonsynonymous site to synonymous substitutions per synonymous site) for Sciaphila (indicated below as "MHT," an abbreviation for "mycoheterotroph"), compared with photosynthetic (green) outgroups.
The corresponding unilateral moves to each state are indicated below the ranking of states for each DM in Table 14.
In this and all experiments indicated below, stimulations were carried out for 24 h.
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