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Automated and multiplexed soft lithography for the production of low-density DNA microarrays

机译:自动化和多重软光刻技术,用于生产低密度DNA微阵列

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摘要

Microarrays are established research tools for genotyping, expression profiling, or molecular diagnostics in which DNA molecules are precisely addressed to the surface of a solid support. This study assesses the fabrication of low-density oligonucleotide arrays using an automated microcontact printing device, the InnoStamp 40(®). This automate allows a multiplexed deposition of oligoprobes on a functionalized surface by the use of a MacroStamp(TM) bearing 64 individual pillars each mounted with 50 circular micropatterns (spots) of 160 µm diameter at 320 µm pitch. Reliability and reuse of the MacroStamp(TM) were shown to be fast and robust by a simple washing step in 96% ethanol. The low-density microarrays printed on either epoxysilane or dendrimer-functionalized slides (DendriSlides) showed excellent hybridization response with complementary sequences at unusual low probe and target concentrations, since the actual probe density immobilized by this technology was at least 10-fold lower than with the conventional mechanical spotting. In addition, we found a comparable hybridization response in terms of fluorescence intensity between spotted and printed oligoarrays with a 1 nM complementary target by using a 50-fold lower probe concentration to produce the oligoarrays by the microcontact printing method. Taken together, our results lend support to the potential development of this multiplexed microcontact printing technology employing soft lithography as an alternative, cost-competitive tool for fabrication of low-density DNA microarrays.
机译:微阵列是用于基因分型,表达谱分析或分子诊断的成熟研究工具,其中将DNA分子精确定位到固体支持物的表面。这项研究评估了使用自动微接触印刷设备InnoStamp40®的低密度寡核苷酸阵列的制造。通过使用带有64个单独的柱子的MacroStamp™,每个柱子都安装有50个直径为160 µm的圆形微图案(点),间距为320 µm,从而实现了自动化,从而在功能化表面上进行寡聚探针的多重沉积。通过在96%乙醇中进行简单的洗涤步骤,显示了MacroStamp™的可靠性和重复使用性快速而稳定。印在环氧硅烷或树枝状大分子官能化载玻片(DendriSlides)上的低密度微阵列在极低的探针和靶标浓度下显示出与互补序列的出色杂交反应,因为通过该技术固定的实际探针密度至少比使用该技术固定的探针密度低10倍。传统的机械点样。此外,我们发现通过使用微接触印刷方法使用低50倍探针浓度生产寡阵列,在具有1 nM互补靶点的点样和印刷寡阵列之间的荧光强度方面,可比的杂交反应。综上所述,我们的结果为这种多重微接触印刷技术的潜在发展提供了支持,该技术采用软光刻技术作为制造低密度DNA微阵列的另一种具有成本竞争力的工具。

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