In a comparison between microarray and NGS technologies in synthetic pools of small RNA, Willenbrock et al. [ 13] found that multiplexing resulted in decreased sensitivity due to a reduction of sequencing depth and a loss of reproducibility; however the authors did not investigate power for detection of DE in their study.
The general in vitro selection procedure for an aptamer, SELEX (Fig. 15), is as follows: a synthetic DNA pool is prepared by chemical synthesis.
Ex-vivo ELISpot assays used Vespula species wasp venom (Vespa Labs, Spring Mills, PA, USA) at 5 µg/ml and also synthetic peptide pools.
The cells were diluted at 2.5×106 PBMC/ml in CTL Test Media Cellular Technology Ltd ... 250 000 PBMCs/well were stimulated in ELISPOT-plates in the presence of synthetic peptide-pool (10 µg/ml each), PPD (10 µg/ml), rMtb-HBHA (25 µg/ml).
In the short-term assay we stimulated PBMC for 16 h with different recombinant M. tuberculosis proteins (5 µg/ml, see Table 1), purified protein derivative (PPD) of M. tuberculosis (5 µg/ml) (Statens Serum Institute), Staphylococcus enterotoxin B (SEB) (1 µg/ml) (Sigma-Aldrich), and synthetic peptide pools (5 µg/ml per peptide) at 37°C and 5% CO2.
Assays of in vitro interferon (IFN -γ secretIFN -γ resecretion recombinant responsend overlapping syntoetic peptide pools frecombinant gRv1737of immunodominand proverlapping performed.
The gp100 protein was spanned by 163 synthetic peptides (16 pools), tyrosinase by 130 peptides (13 pools), MAGE-C2 by 91 peptides (9 pools), and MAGE-A3 by 76 peptides (8 pools) (all purchased from EMC Microcultures, Tübingen, Germany).
We established a chiral pool synthetic route to yield the desired tricyclic chiral isomers.
One intriguing idea, for example, are pooled synthetic lethality screens [ 27, 28] allowing the identification of cancer specific molecular targets.
The antigenic specificity of CD8+ T-cell lines was examined using pooled synthetic peptides (16 mer overlapping by 12 residues) covering the full length of five defined T. parva antigens (Tp1, Tp2, Tp4, Tp5 and Tp8) and peptides representing three defined peptides in Tp1 and Tp2 (Pepscan Systems, Lelystad, The Netherlands).
