Competitive (intermolecular) KIEs were measured using an equimolar mixture of d0- and d23-lauric acid.
Specific activity was quantitatively estimated by measuring the hydrolysis of polygalacturonic acid using an equimolar quantity of AnPGI.
Alternatively, all strains used in microarray design can be used as control if an equimolar mix of their genomic DNA is labeled with the reference fluorochrome and hybridized together with the test.
The microarray hybridisation was performed using a reference design, using a reference RNA sample, which comprised an equimolar mix of RNA extracted from all individual fish and tissue samples.
PCR products were quantified using a Nanodrop 1000 and pooled in an equimolar mix.
The CGH was performed using genomic DNA from the tumor samples co-hybridized with an equimolar mix of 10 ethnically-matched (finnish) DNA samples on NimbleGen 12x135K CGH arrays, which 60-mer probes spaced every 13 kb on average.
A DNA bank enriched in microsatellite markers was developed from an equimolar mix of genomic DNA extracts from four species, including A. bipindensis, using the 454 GS-FLX platform (454 Life Sciences, a Roche Company, Branford, Connecticut, USA) according to the protocols of Malausa et al. (2011) and Micheneau et al. (2011).
One common fluorescent-labelled primer (5'-FAM AGC AAC ATT CAT AGT CTT TG) and the poly A-specific primers 5'-CCA CAC TAA TTG AGC TAA GC-3' (long transcript) and an equimolar mix of 5'-(T)15GandA-3' and 5'-(T)15GGGAA-3' (short transcript) were used to simultaneously amplify two different RT PCR products with length 352 bp and 422 bp, respectively.
The PCR was performed using 5 U of Platinum Taq (Invitrogen), 1x PCR Buffer, 0.5 mM dNTPs, 0.5 μM of Adapter Primer1and 0.5 μM of an equimolar mix of the IS specific primers, 2 mM MgCl2, 0.5 M Betaine and 120 ng (equivalent to 5 ng per sample) of the purified DNA in a final volume of 25 μl.
We used two of these strains as tests (R and G) and hybridized them with two different controls: a single strain (T) or an equimolar mix of the three strains represented in the array (T+R+G).
