High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers
We have developed a new method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The technique involves PCR amplification of genomic DNA with two tailed allele-specific primers that introduce priming sites for universal energy-transfer-labeled primers. The output of red and g...
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| Veröffentlicht in: | Genome research 2001-01, Vol.11 (1), p.163-169 |
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| creator | Myakishev, M V Khripin, Y Hu, S Hamer, D H |
| description | We have developed a new method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The technique involves PCR amplification of genomic DNA with two tailed allele-specific primers that introduce priming sites for universal energy-transfer-labeled primers. The output of red and green light is conveniently scored using a fluorescence plate reader. The new method, which was validated on nine model SNPs, is well suited for high-throughput, automated genotyping because it requires only one reaction per SNP, it is performed in a single tube with no post-PCR handling, the same energy-transfer-labeled primers are used for all analyses, and the instrumentation is inexpensive. Possible applications include multiple-candidate gene analysis, genomewide scans, and medical diagnostics. |
| doi_str_mv | 10.1101/gr.157901 |
| format | Article |
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The technique involves PCR amplification of genomic DNA with two tailed allele-specific primers that introduce priming sites for universal energy-transfer-labeled primers. The output of red and green light is conveniently scored using a fluorescence plate reader. The new method, which was validated on nine model SNPs, is well suited for high-throughput, automated genotyping because it requires only one reaction per SNP, it is performed in a single tube with no post-PCR handling, the same energy-transfer-labeled primers are used for all analyses, and the instrumentation is inexpensive. Possible applications include multiple-candidate gene analysis, genomewide scans, and medical diagnostics.</description><subject>Alleles</subject><subject>DNA Primers - chemical synthesis</subject><subject>DNA Primers - genetics</subject><subject>Energy Transfer - genetics</subject><subject>Fluorescein</subject><subject>Fluorescent Dyes</subject><subject>Genotype</subject><subject>Methods</subject><subject>Polymerase Chain Reaction - instrumentation</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymorphism, Single Nucleotide - genetics</subject><subject>Sensitivity and Specificity</subject><subject>Xanthenes</subject><issn>1088-9051</issn><issn>1549-5469</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkctr3DAQxkVp6ObRQ_-BoFOhByV6rGzp0ENZmmwhJCGPs5Dlka2gtV3JTtj_Pl526eM0A_P7Zj7mQ-gLoxeMUXbZpAsmS03ZB3TM5FITuSz0x7mnShFNJVugk5xfKKViqdQntGCMyaLg8hjV69C0ZGxTPzXtMI348fYeN9D143YIXYOrLbYxQgSSB3DBB4fvVw_4LYwtnrrwCinbiKGD1GzJmGyXPSQSbTVLajyksJmJM3Tkbczw-VBP0fPVz6fVmtzcXf9a_bghTpR0JJJ5cNbVvNAcmJWWc7WUvFJeOed9AbQsZSGop1rXNfhKc60KqLkVmhecilP0fb93mKoN1A662VE0Oxc2bU1vg_l_0oXWNP2rEfMbhZj1Xw_61P-eII9mE7KDGG0H_ZRNSaUqRaln8NsedKnPOYH_c4NRs4vENMnsI5nZ839N_SUPGYh3pAyJsQ</recordid><startdate>200101</startdate><enddate>200101</enddate><creator>Myakishev, M V</creator><creator>Khripin, Y</creator><creator>Hu, S</creator><creator>Hamer, D H</creator><general>Cold Spring Harbor Laboratory Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>200101</creationdate><title>High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers</title><author>Myakishev, M V ; Khripin, Y ; Hu, S ; Hamer, D H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c370t-51fecacd2692e1a5a228452b8f8ccff6e0775630f099ddefb92986ed2a3926203</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Alleles</topic><topic>DNA Primers - chemical synthesis</topic><topic>DNA Primers - genetics</topic><topic>Energy Transfer - genetics</topic><topic>Fluorescein</topic><topic>Fluorescent Dyes</topic><topic>Genotype</topic><topic>Methods</topic><topic>Polymerase Chain Reaction - instrumentation</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polymorphism, Single Nucleotide - genetics</topic><topic>Sensitivity and Specificity</topic><topic>Xanthenes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Myakishev, M V</creatorcontrib><creatorcontrib>Khripin, Y</creatorcontrib><creatorcontrib>Hu, S</creatorcontrib><creatorcontrib>Hamer, D H</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Genome research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Myakishev, M V</au><au>Khripin, Y</au><au>Hu, S</au><au>Hamer, D H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers</atitle><jtitle>Genome research</jtitle><addtitle>Genome Res</addtitle><date>2001-01</date><risdate>2001</risdate><volume>11</volume><issue>1</issue><spage>163</spage><epage>169</epage><pages>163-169</pages><issn>1088-9051</issn><eissn>1549-5469</eissn><abstract>We have developed a new method for high-throughput genotyping of single nucleotide polymorphisms (SNPs). 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| ispartof | Genome research, 2001-01, Vol.11 (1), p.163-169 |
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| subjects | Alleles DNA Primers - chemical synthesis DNA Primers - genetics Energy Transfer - genetics Fluorescein Fluorescent Dyes Genotype Methods Polymerase Chain Reaction - instrumentation Polymerase Chain Reaction - methods Polymorphism, Single Nucleotide - genetics Sensitivity and Specificity Xanthenes |
| title | High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers |
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