Capillary-based fully integrated and automated system for nanoliter polymerase chain reaction analysis directly from cheek cells
Capillary-based fully integrated and automated system for nanoliter polymerase chain reaction analysis directly from cheek cells
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DOI：
10.1016/s0021-9673(01)00828-7
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发表时间：
2001-07-27
影响因子：
4.1
通讯作者：
Yeung, ES
中科院分区：
文献类型：
--
作者：
He, Y;Zhang, YHH;Yeung, ES
一个小型化，集成化和自动化的系统的基础上毛细管射流已经开发出纳升DNA分析直接从脸颊细胞。DNA分析的所有步骤，包括注入水性试剂和DNA样品，将溶液混合在一起，热细胞裂解，聚合酶链反应（PCR），PCR产物的转移和注入，这些产物的分离，大小测定和检测都在基于毛细管的集成系统中进行。将通过塑料牙签收集的少量颊细胞直接溶解在塑料小瓶中的PCR混合物中，或在毛细管中与小体积的PCR混合物（125 nl）在线混合。在微热循环仪中进行热细胞裂解和PCR后，将DNA片段与DNA大小标准品混合，并转移至微交叉进样，通过毛细管凝胶电泳进行分离。可编程注射泵、开关阀、多位阀和冻融阀用于整个系统中的微流体控制。这项工作确立了在基于毛细管的纳升集成系统中从真实的样品进行DNA分析的所有步骤的可行性。(C)2001 Elsevier Science B.V.保留所有权利。
A miniaturized, integrated and automated system based on capillary fluidics has been developed for nanoliter DNA analysis directly from cheek cells. All steps for DNA analysis, including injecting aqueous reagents and DNA samples, mixing the solutions together, thermal cell lysis, polymerase chain reaction (PCR), transfer and injection of PCR product, separation, sizing and detection of those products are performed in a capillary-based integrated system. A small amount of cheek cells collected by a plastic toothpick is directly dissolved in the PCR cocktail in a plastic vial or mixed on-line with a small volume of PCR cocktail (125 nl) in the capillary. After thermal cell lysis and PCR in a microthermal cycler, the DNA fragments are mixed with DNA size standards and transferred to a micro-cross for injection and separation by capillary gel electrophoresis. Programmable syringe pumps, switching valves, multiposition and freeze-thaw valves are used for microfluidic control in the entire system. This work establishes the feasibility of performing all the steps of DNA analysis from real samples in a capillary-based nanoliter integrated system. (C) 2001 Elsevier Science B.V. All rights reserved.