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Pooled reactions were genotyped using an ABI 3730 DNA Genetic Analyzer and visualised using Genescan.
The second approach, referred to as the pooled reactions consortium analysis, treated the consortium as a single entity.
The pooled reactions consortium analysis method modeled community metabolic potential by treating all enzymatic activities and metabolites as residents of the same physical space, without membrane boundaries.
An additional advantage of the pooled reactions approach is the reduction of computational burden.
The pooled reactions approach represents the coarsest-scale methodology, requires the least a priori information, and is easier to implement than alternative approaches.
First, pooled reactions can result in higher predicted yields than compartmentalized reactions because there are no logistical costs associated with inter-guild metabolite transfer.
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PCR reactions were cleaned up with AMPure XP (Beckman Coulter Genomics), and pooled reaction yields were quantified using the Qubit fluorometer (Invitrogen).
Authors' response: Yes, I wish I had thought of that at the time (i.e., pooling reactions using smaller numbers of cycles).
We compared reactions inside and outside the range of venomous snakes using a two-tailed Fisher's exact test, pooling reactions to venomous and non-venomous snakes inside the range of venomous snakes due to the small total sample size.
Many excitation processes such as electron collision excitation from the molecular ground state ( {text{N}}_{2} ({text{X}}^{1} varSigma_{g}^ ) ), the first metastable state ( {text{N}}_{2} ({text{A}}^{3} varSigma_{u}^ ) ), pooling reaction and transfer of energy between collisional partners would result in these transitions [27, 28].
Therefore, we processed at least three replicate 10,000 cell reactions for each sonication cycle number tested, and pooled the reactions prior to the DNA recovery steps.
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