Department of Chemical Biology
Yang's Lab,Department of Chemical Biology, Xiamen University
Professor Yang
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Professor
Department of Chemical Biology, Xiamen University
Contact Information
Yang's LaboratoryRoom 532, Lujiaxi Building, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China
Ph: +86 (0) 592-218 7601cyyang@xmu.edu.cn
Fenxiang's work on Palmtop Sequencer has been accepted by Biosens Bioelectron
2018-09-28 12:12:11
To explore genome mutation meaningfully, it is in urgent need to develop an automated and inexpensive platform for DNA mutations analysis. Digital microfluidics (DMF) is a powerful platform for a broad range of applications due to the advantages of high automatization and low reagent consumption. Pyrosequencing enables DNA sequencing based on non-electrophoresis bioluminescence, which is suitable for rapid and sensitive analysis of short sequences. Herein, we describe a palmtop sequencing platform for automatic, real-time and portable analysis of DNA mutations, which is based on the pyrosequencing principle and implemented by digital microfluidics. The portable system can sequence a DNA template with up to 53 bp with 100% accuracy within 2 hours. Mutation in the KRAS gene can be detected within 30 minutes with a LOD as low as 5% mutant level. Portable and accurate gender identification was further demonstrated by sequencing a short amelogenin (AMEL) fragment. With the advantages of portability, ease of use, high accuracy, andlow cost, the palmtop sequencing platform shows great potential for portable genetic testing in a variety of circumstances.
基于其离散液滴精准控制和自动化的优点，课题组在数字微流控平台上结合焦磷酸测序发展了快速实时自动化的基因突变检测装置---掌上测序仪。该测序仪包括操纵液滴的数字微流控系统、进行磁珠分离的磁模块和化学发光采集模块。在实验中，将焦磷酸测序试剂（包括样品、酶和dNTP）分别置于芯片的储液槽中，通过软件编程系统操控液滴的移动，实现焦磷酸测序实验中所需的加样、洗涤、分离等一系列繁琐的操作。
研究成员将其应用于短序列的突变检测以及单核苷酸多态性位点（SNP）分析中。通过掌上焦磷酸测序平台进行KRAS基因外显子2第12和13密码子突变的检测，在检测中，掌上测序仪能够在30分钟内检测出低至5%的突变水平。通过掌上焦磷酸测序仪检测短片段牙釉质蛋白基因上的单核苷酸多态性位点，能够准确进行性别的判定。实验结果表明，该平台在多步骤生物化学的实际应用中具有很高的潜力，可用于多种应用如SNP基因分型、微生物鉴定、肿瘤突变检测以及定量CpG岛甲基化分析。
基于其离散液滴精准控制和自动化的优点，课题组在数字微流控平台上结合焦磷酸测序发展了快速实时自动化的基因突变检测装置---掌上测序仪。该测序仪包括操纵液滴的数字微流控系统、进行磁珠分离的磁模块和化学发光采集模块。在实验中，将焦磷酸测序试剂（包括样品、酶和dNTP）分别置于芯片的储液槽中，通过软件编程系统操控液滴的移动，实现焦磷酸测序实验中所需的加样、洗涤、分离等一系列繁琐的操作。
研究成员将其应用于短序列的突变检测以及单核苷酸多态性位点（SNP）分析中。通过掌上焦磷酸测序平台进行KRAS基因外显子2第12和13密码子突变的检测，在检测中，掌上测序仪能够在30分钟内检测出低至5%的突变水平。通过掌上焦磷酸测序仪检测短片段牙釉质蛋白基因上的单核苷酸多态性位点，能够准确进行性别的判定。实验结果表明，该平台在多步骤生物化学的实际应用中具有很高的潜力，可用于多种应用如SNP基因分型、微生物鉴定、肿瘤突变检测以及定量CpG岛甲基化分析。