Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells.
Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells.
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DOI：
10.1039/c2lc40955k
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发表时间：
2012-11-21
期刊：
影响因子：
6.1
通讯作者：
Craighead HG
中科院分区：
文献类型：
--
作者：
Benítez JJ;Topolancik J;Tian HC;Wallin CB;Latulippe DR;Szeto K;Murphy PJ;Cipriany BR;Levy SL;Soloway PD;Craighead HG
我们描述了一种用于从单细胞中提取、纯化和拉伸人类染色体DNA的微流控设备。微流控聚二甲基硅氧烷通道中的二维微柱阵列被设计用于捕获单个人体细胞。裂解后从细胞中释放出的百万碱基长的DNA链被捕获在微柱阵列中，并在最佳的流体动力流动条件下拉伸。完整的染色体DNA被缠绕在阵列中，而其他细胞成分被从通道中洗掉。为了演示捕获原理，将单个染色体与整个染色体颜料杂交，并在荧光显微镜下成像。从单个细胞和小细胞群(小于100个)中提取的DNA在连续流动下通过限制性内切酶消化从设备中释放出来，并收集用于芯片外分析。对提取的材料进行量化表明，该微设备有效地提取了几乎所有的染色体DNA。所描述的装置代表了一种在单细胞水平上对染色体DNA进行各种分析的新平台。
We describe a microfluidic device for the extraction, purification and stretching of human chromosomal DNA from single cells. A two-dimensional array of micropillars in a microfluidic polydimethylsiloxane channel was designed to capture a single human cell. Megabase-long DNA strands released from the cell upon lysis are trapped in the micropillar array and stretched under optimal hydrodynamic flow conditions. Intact chromosomal DNA is entangled in the array, while other cellular components are washed from the channel. To demonstrate the entrapment principle, a single chromosome was hybridized to whole chromosome paints, and imaged by fluorescence microscopy. DNA extracted from a single cell and small cell populations (less than 100) was released from the device by restriction endonuclease digestion under continuous flow and collected for offchip analysis. Quantification of the extracted material reveals that the microdevice efficiently extracts essentially all chromosomal DNA. The device described represents a novel platform to perform a variety of analyses on chromosomal DNA at the single cell level.
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影响因子：
6.1
作者：
Reedy CR;Price CW;Sniegowski J;Ferrance JP;Begley M;Landers JP
通讯作者：
Landers JP
影响因子：
4.5
作者：
Marcy, Yann;Ishoey, Thomas;Lasken, Roger S.;Stockwell, Timothy B.;Walenz, Brian P.;Halpern, Aaron L.;Beeson, Karen Y.;Goldberg, Susanne M. D.;Quake, Stephen R.
通讯作者：
Quake, Stephen R.
影响因子：
64.8
作者：
Navin N;Kendall J;Troge J;Andrews P;Rodgers L;McIndoo J;Cook K;Stepansky A;Levy D;Esposito D;Muthuswamy L;Krasnitz A;McCombie WR;Hicks J;Wigler M
通讯作者：
Wigler M
影响因子：
1.4
作者：
Turner, SW;Perez, AM;Craighead, HG
通讯作者：
Craighead, HG
影响因子：
64.8
作者：
VOLKMUTH, WD;AUSTIN, RH
通讯作者：
AUSTIN, RH