Subsequently, litter was pooled according to species and origin of leaf litter, i.e., urban and periurban tree locations, yielding ten pools in total.
This gave 30 pools in total (5 families × 3 BT × 2 tails (high and low)).
All 5 sampling dates could be completed, yielding a sample of 187 pools in total, equivalent to 935 individual fecal pellets.
Pools of retroviral vectors encoding 354 human kinases and kinase-related proteins (37 pools in total, each consisting of 10 12 unique open reading frames) were transduced into HMED-DD-NeuT cells.
Thus, 14 and 4 proteins were identified by MS from the protein spots recognised by at least 75% of the 15 pools in total and enriched nuclear protein extracts, respectively (Table 2).
Briefly, the BACs were pooled from 42 384-well plates (i.e. 384 BACs with a 138 kb average insert size which per plate covers ~53 Mb (23%) of the 230 Mb), in a 3-D fashion into row, column and splitbox pools resulting in 22 pools per plate (924 pools in total).
In total, permutations of the original 18 reference-sample sequence pairs (3× replicates at 3× GC contents and 2× divergence levels, with 3× read length distributions, 3× damage levels and 7× coverage depths) presented 378 sets of triplicate read pools, or 1134 read pools in total.
Fourteen and four proteins were recognised by IgG from at least 75% of the 15 pools in total and enriched nuclear protein extracts, respectively, whereas 15 protein spots were specifically recognised by IgG from at least four of the ten pools from patients with unidentified ANAs.
Finally, among the spots recognised by IgG from the 10 pools of sera from patients with unidentified ANAs, 15 were specifically recognised by IgG from at least 4 of these 10 pools in total or enriched nuclear protein extracts (Table 3) aSSc: systemic sclerosis.
