Nanoliter multiplex PCR arrays on a SlipChip.
Nanoliter multiplex PCR arrays on a SlipChip.
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DOI：
10.1021/ac1007249
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发表时间：
2010-06-01
影响因子：
7.4
通讯作者：
Ismagilov, Rustem F.
中科院分区：
文献类型：
--
作者：
Shen, Feng;Du, Wenbin;Davydova, Elena K.;Karymov, Mikhail A.;Pandey, Janmajay;Ismagilov, Rustem F.
对SlipChip平台进行了高通量纳升多重聚合酶链式反应的测试。使用SlipChip平台进行多重聚合酶链式反应的优势包括：能够预加载干引物阵列、无需仪器操作样品、样品体积小、吞吐能力高。SlipChip的设计是在每个反应隔间预装一对引物，并在每个反应隔室筛选多达384对不同的引物对，样品不超过30纳升。SlipChip的40孔和384孔设计都被测试了多重PCR。在这里使用的几何形状中，甚至在SlipChip的两个板滑动之前，样品流体就自发地被分割成不同的体积，但滑动引入了额外的功能，使设备更加坚固和多功能。SlipChip的井是为了克服与热膨胀相关的潜在问题而设计的。通过使用充满油的圆形井，并将它们与填充有PCR水混合物的方形井重叠，PCR水混合物液滴总是被润滑液包围。在这种设计中，在加热和热膨胀过程中，只有油从隔室中排出，防止了水溶液的泄漏。发现40孔和384孔装置都没有交叉污染，终点荧光检测提供了可靠的读数。多个样品也可以在同一个SlipChip上同时进行筛选。在384孔芯片上用20对不同的引物进行多重聚合酶链式反应，以鉴定血液感染中常见的16种细菌和真菌。SlipChip在单独的实验中正确地识别了五种不同的细菌或真菌。此外，还鉴定了耐甲氧西林金黄色葡萄球菌(MRSA)中存在耐药基因mecA。SlipChip将用于涉及聚合酶链式反应阵列的应用，并为诊断、护理点设备和基于固定的阵列的新战略奠定基础。
The SlipChip platform was tested to perform high throughput nanoliter multiplex PCR. The advantages of using the SlipChip platform for multiplex PCR include the ability to preload arrays of dry primers, instrument-free sample manipulation, small sample volume, and high throughput capacity. The SlipChip was designed to preload one primer pair per reaction compartment, and to screen up to 384 different primer pairs with less than 30 nanoliters of sample per reaction compartment. Both a 40-well and a 384-well design of the SlipChip were tested for multiplex PCR. In the geometries used here, the sample fluid was spontaneously compartmentalized into discrete volumes even before slipping of the two plates of the SlipChip, but slipping introduced additional capabilities that made devices more robust and versatile. The wells of this SlipChip were designed to overcome potential problems associated with thermal expansion. By using circular wells filled with oil and overlapping them with square wells filled with the aqueous PCR mixture, a droplet of aqueous PCR mixture was always surrounded by the lubricating fluid. In this design, during heating and thermal expansion, only oil was expelled from the compartment and leaking of the aqueous solution was prevented. Both 40-well and 384-well devices were found to be free from cross-contamination, and end point fluorescence detection provided reliable readout. Multiple samples could also be screened on the same SlipChip simultaneously. Multiplex PCR was validated on the 384-well SlipChip with 20 different primer pairs to identify 16 bacterial and fungal species commonly presented in blood infections. The SlipChip correctly identified five different bacterial or fungal species in separate experiments. In addition, the presence of the resistance gene mecA in methicillin resistant Staphylococcus aureus (MRSA) was identified. The SlipChip will be useful for applications involving PCR arrays, and lays the foundation for new strategies for diagnostics, point-of-care devices, and immobilization based arrays.
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前列腺癌的基因表达谱揭示了多个分子途径在转移过程中的参与。
影响因子：
3.8
作者：
Chandran, Uma R.;Ma, Changqing;Dhir, Rajiv;Bisceglia, Michelle;Lyons-Weiler, Maureen;Liang, Wenjing;Michalopoulos, George;Becich, Michael;Monzon, Federico A.
通讯作者：
Monzon, Federico A.
影响因子：
15
作者：
Kreutz JE;Li L;Roach LS;Hatakeyama T;Ismagilov RF
通讯作者：
Ismagilov RF
DOI：
10.1073/pnas.88.23.10921
发表时间：
1991-12-01
影响因子：
11.1
作者：
HOWELL, MD;DIVELEY, JP;BROSTOFF, SW
通讯作者：
BROSTOFF, SW
影响因子：
3.5
作者：
Bustin, SA
通讯作者：
Bustin, SA
影响因子：
6.7
作者：
Aryan, Ehsan;Makvandi, Manoochehr;Romano, Marta
通讯作者：
Romano, Marta