Microarrays were made of 389,307 spots of 50-nt probes.
Microarrays are made up of thousands to millions of microscopic "features", clusters of identical oligonucleotide probes, which are used to detect hybridized gene products.
Microarrays were made by spotting purified ds-PCR products 500 to 1200 bp in length onto glass slides coated with aminopropylsilane (Erie Scientific, Portsmouth, New Hampshire, USA).
The single-stranded DNA 60-mer ETS↔CRE microarrays were made double-stranded by primer extension as described previously (Badis et al. 2009).
Both the design of 70-mer oligonuclearotides for each gene and the construction of microarrays were made by CapitalBio Corporation (Beijing, China).
Microarrays can be made by presynthesizing the probe molecule and spotting it on a surface using appropriate tethering chemistry, but modern microarrays are made with in situ methods in which the biomolecules are synthesized directly on the substrate from their monomer components, which allows for high probe densities, high uniformity, and high reproducibility.
Similar conclusions on hybridization kinetics using conventional microarrays were made by Pappaert et al.; they showed that a shear-driven flow reduces the analysis time (from 16 hours down to 30 minutes) [ 79].
The first high-density microarrays were made by spotting large numbers of individual complementary DNA (cDNA) molecules, cDNA reverse transcribed from mRNA, as probes onto polylysinated glass slides in an arrayed pattern where the hybridized targets could be identified by the position of the probe [ 5].
This is demonstrated in Figure 1, where microarrays were made by printing the Gal4-pZsGreen reporter mixed with pBD-p53 and pAD-SV40T, coding for two interacting proteins or mixed with pBD-p53 and pAD-TRAF, coding for two non-interacting proteins.
The microarray was made as previously described (Huang et al., 2006).
