Recovery of genomic DNA from residual frozen archival blood clots suitable for amplification and use in genotyping assays
被引:14
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作者:
Iovannisci, DM
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机构:Res Ctr Oakland, Oakland, CA USA
Iovannisci, DM
Ha, TT
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机构:Res Ctr Oakland, Oakland, CA USA
Ha, TT
Shaw, GM
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机构:Res Ctr Oakland, Oakland, CA USA
Shaw, GM
机构:
[1] Res Ctr Oakland, Oakland, CA USA
[2] Calif Birth Defects Monitoring Program, March Dimes Birth Defects Fdn, Berkeley, CA USA
来源:
GENETIC TESTING
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2006年
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10卷
/
01期
关键词:
D O I:
10.1089/gte.2006.10.44
中图分类号:
Q3 [遗传学];
学科分类号:
071007 ;
090102 ;
摘要:
A greatly neglected source of DNA potentially useful for genetic or forensic studies is the clot remaining from blood samples collected for serum chemistry measurements. We have investigated the utility of residual clots remaining from venipunctures collected for California's Expanded Maternal Serum Alpha-Fetoprotein Screening Program. We report a protocol based on the salting out method for the extraction of DNA from samples which have been archived and frozen for up to 2.5 years. As much as 57 mu g of high-quality DNA can be obtained from a 2-ml clot as determined by PicoGreen (R) ( Molecular Probes, Inc., Eugene, OR) fluorescence measurements. Quality of the purified DNA was evaluated by its ability to serve as template in polymerase chain reaction (PCR) amplifications, using primers that flank the polymorphic regions of six genes of pharmacogenetic interest distributed throughout the human genome. Sizes of the gene regions successfully amplified range from 215 bp to 2064 bp, using as little as 10 ng of template DNA. Because many genotyping protocols routinely recommend the design of amplicons in the 100-200 bp range, and 10-50 ng of template, we conclude that the clot remaining after serum has been removed from blood collected for serum chemistry measurements can serve as a reliable source of DNA for genotyping studies.
机构:
Stanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
Stanford Univ, Howard Hughes Med Inst, Sch Med, Stanford, CA 94305 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
St Julien, Krystal R.
Jelliffe-Pawlowski, Laura L.
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机构:
Calif Dept Publ Hlth, Calif Genet Dis Screening Program, Richmond, VA USA
Univ Calif San Francisco, Dept Epidemiol & Biostat, San Francisco, CA 94143 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
Jelliffe-Pawlowski, Laura L.
Shaw, Gary M.
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机构:
Stanford Univ, Dept Pediat, Sch Med, Stanford, CA 94305 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
Shaw, Gary M.
Stevenson, David K.
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机构:
Stanford Univ, Dept Pediat, Sch Med, Stanford, CA 94305 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
Stevenson, David K.
O'Brodovich, Hugh M.
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机构:
Stanford Univ, Dept Pediat, Sch Med, Stanford, CA 94305 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
O'Brodovich, Hugh M.
Krasnow, Mark A.
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机构:
Stanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
Stanford Univ, Howard Hughes Med Inst, Sch Med, Stanford, CA 94305 USAStanford Univ, Dept Biochem, Sch Med, Stanford, CA 94305 USA
机构:
Brown Univ, Ctr Biomed Engn, Sch Engn, Providence, RI USA
Brown Univ Sch Publ Hlth, Sch Publ Hlth, Providence, RI USABrown Univ, Ctr Biomed Engn, Sch Engn, Providence, RI USA
Lee, Kiara
Tripathi, Anubhav
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机构:
Brown Univ, Ctr Biomed Engn, Sch Engn, Providence, RI USA
Brown Univ, Ctr Biomed Engn, Sch Engn, Providence, RI 02912 USABrown Univ, Ctr Biomed Engn, Sch Engn, Providence, RI USA
机构:
ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Vikrama Simhapuri Univ, Dept Biotechnol, Nellore 524003, Andhra Pradesh, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Goud, Talla Sridhar
Upadhyay, Ramesh Chandra
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ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Upadhyay, Ramesh Chandra
Kumar, Anil
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机构:
ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Kumar, Anil
Karri, Satyanagalakshmi
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机构:
Vikrama Simhapuri Univ, Dept Biotechnol, Nellore 524003, Andhra Pradesh, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Karri, Satyanagalakshmi
Choudhary, Renuka
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ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Choudhary, Renuka
Ashraf, Syma
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机构:
ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Ashraf, Syma
Singh, Sohan Vir
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机构:
ICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Singh, Sohan Vir
Kumar, Onteru Suneel
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机构:
ICAR Natl Dairy Res Inst, Anim Biochem Div, Mol Endocrinol & Struct Biol, Karnal 132001, Haryana, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India
Kumar, Onteru Suneel
Kiranmai, Chadipiralla
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机构:
Vikrama Simhapuri Univ, Dept Biotechnol, Nellore 524003, Andhra Pradesh, IndiaICAR Natl Dairy Res Inst, Climate Resilient Live Stock Res Ctr, Karnal 132001, Haryana, India