Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform
This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD 15/45 antibodies,...
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| Veröffentlicht in: | Microfluidics and nanofluidics 2009-04, Vol.6 (4), p.539-555 |
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| creator | Lien, Kang-Yi Liu, Chien-Ju Lin, Yi-Chien Kuo, Pao-Lin Lee, Gwo-Bin |
| description | This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD
15/45
antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene. |
| doi_str_mv | 10.1007/s10404-008-0337-x |
| format | Article |
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15/45
antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.</description><identifier>ISSN: 1613-4982</identifier><identifier>EISSN: 1613-4990</identifier><identifier>DOI: 10.1007/s10404-008-0337-x</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>Analytical Chemistry ; Biological and medical sciences ; Biomedical Engineering and Bioengineering ; Deoxyribonucleic acid ; Diverse techniques ; DNA ; Engineering ; Engineering Fluid Dynamics ; Fundamental and applied biological sciences. Psychology ; Genetic disorders ; Molecular and cellular biology ; Nanotechnology and Microengineering ; Nucleic acids ; Research Paper</subject><ispartof>Microfluidics and nanofluidics, 2009-04, Vol.6 (4), p.539-555</ispartof><rights>Springer-Verlag 2008</rights><rights>2009 INIST-CNRS</rights><rights>Springer-Verlag 2009</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c378t-e2a5babac3b32403cb52499bace5113d8144a517308b7b81479ac09d32f155543</citedby><cites>FETCH-LOGICAL-c378t-e2a5babac3b32403cb52499bace5113d8144a517308b7b81479ac09d32f155543</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10404-008-0337-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10404-008-0337-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,778,782,27907,27908,41471,42540,51302</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21346045$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Lien, Kang-Yi</creatorcontrib><creatorcontrib>Liu, Chien-Ju</creatorcontrib><creatorcontrib>Lin, Yi-Chien</creatorcontrib><creatorcontrib>Kuo, Pao-Lin</creatorcontrib><creatorcontrib>Lee, Gwo-Bin</creatorcontrib><title>Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform</title><title>Microfluidics and nanofluidics</title><addtitle>Microfluid Nanofluid</addtitle><description>This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD
15/45
antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.</description><subject>Analytical Chemistry</subject><subject>Biological and medical sciences</subject><subject>Biomedical Engineering and Bioengineering</subject><subject>Deoxyribonucleic acid</subject><subject>Diverse techniques</subject><subject>DNA</subject><subject>Engineering</subject><subject>Engineering Fluid Dynamics</subject><subject>Fundamental and applied biological sciences. 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Psychology</topic><topic>Genetic disorders</topic><topic>Molecular and cellular biology</topic><topic>Nanotechnology and Microengineering</topic><topic>Nucleic acids</topic><topic>Research Paper</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lien, Kang-Yi</creatorcontrib><creatorcontrib>Liu, Chien-Ju</creatorcontrib><creatorcontrib>Lin, Yi-Chien</creatorcontrib><creatorcontrib>Kuo, Pao-Lin</creatorcontrib><creatorcontrib>Lee, Gwo-Bin</creatorcontrib><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection (ProQuest)</collection><collection>Natural Science Collection (ProQuest)</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 2: Ocean Technology, Policy & Non-Living Resources</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>ProQuest Engineering Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Engineering Database</collection><collection>Environmental Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>DELNET Engineering & Technology Collection</collection><collection>Nucleic Acids Abstracts</collection><jtitle>Microfluidics and nanofluidics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lien, Kang-Yi</au><au>Liu, Chien-Ju</au><au>Lin, Yi-Chien</au><au>Kuo, Pao-Lin</au><au>Lee, Gwo-Bin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform</atitle><jtitle>Microfluidics and nanofluidics</jtitle><stitle>Microfluid Nanofluid</stitle><date>2009-04-01</date><risdate>2009</risdate><volume>6</volume><issue>4</issue><spage>539</spage><epage>555</epage><pages>539-555</pages><issn>1613-4982</issn><eissn>1613-4990</eissn><abstract>This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD
15/45
antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><doi>10.1007/s10404-008-0337-x</doi><tpages>17</tpages></addata></record> |
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| identifier | ISSN: 1613-4982 |
| ispartof | Microfluidics and nanofluidics, 2009-04, Vol.6 (4), p.539-555 |
| issn | 1613-4982 1613-4990 |
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| source | Springer Nature - Complete Springer Journals |
| subjects | Analytical Chemistry Biological and medical sciences Biomedical Engineering and Bioengineering Deoxyribonucleic acid Diverse techniques DNA Engineering Engineering Fluid Dynamics Fundamental and applied biological sciences. Psychology Genetic disorders Molecular and cellular biology Nanotechnology and Microengineering Nucleic acids Research Paper |
| title | Extraction of genomic DNA and detection of single nucleotide polymorphism genotyping utilizing an integrated magnetic bead-based microfluidic platform |
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