Maskless fabrication of light-directed oligonucleotide microarrays using a digital micromirror array
Maskless fabrication of light-directed oligonucleotide microarrays using a digital micromirror array
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DOI：
10.1038/13664
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发表时间：
1999-10-01
影响因子：
46.9
通讯作者：
Cerrina, F
中科院分区：
文献类型：
--
作者：
Singh-Gasson, S;Green, RD;Cerrina, F
寡核苷酸微阵列，也被称为“DNA芯片”，目前是由光导化学制造的，每个芯片需要大量的光刻掩模。在这里，我们描述了一种无掩模阵列合成器（MAS），它用计算机生成的虚拟掩模取代铬掩模，并将其中继到数字微镜阵列。1:1反射成像系统在玻璃基板的活动表面上形成虚拟掩模的紫外线图像，该玻璃基板安装在与DNA合成器连接的流动池反应室中。程序化的化学偶联循环遵循光照，这些步骤用不同的虚拟掩模重复，以选定的模式生长所需的寡核苷酸。该仪器已用于合成含有超过76,000个特征的寡核苷酸微阵列，测量16 μ m(2)。这些寡核苷酸以高重复产率合成，杂交后可以很容易地区分单碱基对错配。MAS适用于制造包含数千个基因探针的DNA芯片，以及在高密度微阵列中进行的任何其他固相组合化学。
Oligonucleotide microarrays, also called "DNA chips," are currently made by a light-directed chemistry that requires a large number of photolithographic masks for each chip. Here we describe a maskless array synthesizer (MAS) that replaces the chrome masks with virtual masks generated on a computer, which are relayed to a digital micromirror array. A 1:1 reflective imaging system forms an ultraviolet image of the virtual mask on the active surface of the glass substrate, which is mounted in a flow cell reaction chamber connected to a DNA synthesizer. Programmed chemical coupling cycles follow light exposure, and these steps are repeated with different virtual masks to grow desired oligonucleotides in a selected pattern. This instrument has been used to synthesize oligonucleotide microarrays containing more than 76,000 features measuring 16 mu m(2). The oligonucleotides were synthesized at high repetitive yield and, after hybridization, could readily discriminate single-base pair mismatches. The MAS is adaptable to the fabrication of DNA chips containing probes for thousands of genes, as well as any other solid-phase combinatorial chemistry to be performed in high-density microarrays.