Abstract
BackgroundA critical step in the fabrication of biochips is the controlled placement of probes molecules on solid surfaces. This is currently performed by sequential deposition of probes on a target surface with split or solid pins. In this article, we present a cost-effective procedure namely microcontact printing using stamps, for a parallel deposition of probes applicable for manufacturing biochips.ResultsContrary to a previous work, we showed that the stamps tailored with an elastomeric poly(dimethylsiloxane) material did not require any surface modification to be able to adsorb oligonucleotides or PCR products. The adsorbed DNA molecules are subsequently printed efficiently on a target surface with high sub-micron resolution. Secondly, we showed that successive stamping is characterized by an exponential decay of the amount of transferred DNA molecules to the surface up the 4th print, then followed by a second regime of transfer that was dependent on the contact time and which resulted in reduced quality of the features. Thus, while consecutive stamping was possible, this procedure turned out to be less reproducible and more time consuming than simply re-inking the stamps between each print. Thirdly, we showed that the hybridization signals on arrays made by microcontact printing were 5 to 10-times higher than those made by conventional spotting methods. Finally, we demonstrated the validity of this microcontact printing method in manufacturing oligonucleotides arrays for mutations recognition in a yeast gene.ConclusionThe microcontact printing can be considered as a new potential technology platform to pattern DNA microarrays that may have significant advantages over the conventional spotting technologies as it is easy to implement, it uses low cost material to make the stamp, and the arrays made by this technology are 10-times more sensitive in term of hybridization signals than those manufactured by conventional spotting technology.
Highlights
A critical step in the fabrication of biochips is the controlled placement of probes molecules on solid surfaces
The first platform uses the immobilization of prefabricated DNA or oligonucleotides by spotting on functionalized glass slides using metal pins as originally developed by Brown and collaborators, or by a non-contact method using piezoelectric liquid handling [1]
In contrast to this report, we found that untreated PDMS stamp that has a strong hydrophobic surface after curing, adsorbs a sufficient amount of DNA molecules within few seconds while allowing their subsequent deposition by contact on microscope glass slides or silicon
Summary
A critical step in the fabrication of biochips is the controlled placement of probes molecules on solid surfaces. One molecule is deposited after another or one base is added to the previous one, with the consequence that each array is made as an original with a reduced throughput, Affymetrix microarrays manufacturing involves combinatorial processes that allow multiple microarrays (around 96) to be synthesized in parallel in matters of hours. These technology platforms needs sophisticated equipment, leading to high density arrays that can be too expensive for production and utilization of simple-customized-DNA arrays
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