DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors
The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience colu...
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| Veröffentlicht in: | Revista da Sociedade Brasileira de Medicina Tropical 2009-11, Vol.42 (6), p.651-656 |
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| creator | Cardozo, Daniela Maira Guelsin, Gláucia Andréia Clementino, Samaia Laface Melo, Fabiano Cavalcante de Braga, Marco Antônio Souza, Cleonice de Moliterno, Ricardo Alberto Visentainer, Jeane Eliete Laguila |
| description | The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes. |
| doi_str_mv | 10.1590/S0037-86822009000600008 |
| format | Article |
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Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes.</description><identifier>ISSN: 0037-8682</identifier><identifier>EISSN: 1678-9849</identifier><identifier>DOI: 10.1590/S0037-86822009000600008</identifier><identifier>PMID: 20209349</identifier><language>por</language><publisher>Brazil: Sociedade Brasileira de Medicina Tropical</publisher><subject>Blood ; Deoxyribonucleic acid ; DNA ; DNA - blood ; DNA - isolation & purification ; Genotype ; HLA Antigens - genetics ; Humans ; Luminescent Measurements ; Polymerase Chain Reaction - methods ; Receptors, KIR - genetics</subject><ispartof>Revista da Sociedade Brasileira de Medicina Tropical, 2009-11, Vol.42 (6), p.651-656</ispartof><rights>Copyright Sociedade Brasileira de Medicina Tropical Dec 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,865,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20209349$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cardozo, Daniela Maira</creatorcontrib><creatorcontrib>Guelsin, Gláucia Andréia</creatorcontrib><creatorcontrib>Clementino, Samaia Laface</creatorcontrib><creatorcontrib>Melo, Fabiano Cavalcante de</creatorcontrib><creatorcontrib>Braga, Marco Antônio</creatorcontrib><creatorcontrib>Souza, Cleonice de</creatorcontrib><creatorcontrib>Moliterno, Ricardo Alberto</creatorcontrib><creatorcontrib>Visentainer, Jeane Eliete Laguila</creatorcontrib><title>DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors</title><title>Revista da Sociedade Brasileira de Medicina Tropical</title><addtitle>Rev Soc Bras Med Trop</addtitle><description>The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes.</description><subject>Blood</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA - blood</subject><subject>DNA - isolation & purification</subject><subject>Genotype</subject><subject>HLA Antigens - genetics</subject><subject>Humans</subject><subject>Luminescent Measurements</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Receptors, KIR - genetics</subject><issn>0037-8682</issn><issn>1678-9849</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNpd0MtO3TAQBmCrKioH2ldoLXXBKq2vib1EtFwkBAvoOnLsycHg2KljS5zH4I0bLu2ii9G_-Wb0axD6Qsk3KjX5fkMI7xrVKsYI0YSQdh2i3qENbTvVaCX0e7T5h_bRwbLcE8I6rtkHtM8II5oLvUFPP66OMTyWbGzxKeIxpwnbZLY1mAIO39XJRDyElBweU8ZmnoO35sX6iLcQU9nNPm5xAXsX_e8Kywt8XQxQH5LdFcAmFr_qNR3201Rj2oY01OBjE_wD4AwW5pLy8hHtjSYs8OktD9Gv05-3J-fN5fXZxcnxZTPTVpVmEEwpJ51lBojVqqOKGmM6O4DSkis3joy1XAvNpHR8VKCksXZUtNMGBscP0dHr3Tmn59aln_xiIQQTIdWl7ziXSnAhVvn1P3mfao5ruZ4KoamUkpNVfX5TdZjA9XP2k8m7_u-v-R-DuYXi</recordid><startdate>20091101</startdate><enddate>20091101</enddate><creator>Cardozo, Daniela Maira</creator><creator>Guelsin, Gláucia Andréia</creator><creator>Clementino, Samaia Laface</creator><creator>Melo, Fabiano Cavalcante de</creator><creator>Braga, Marco Antônio</creator><creator>Souza, Cleonice de</creator><creator>Moliterno, Ricardo Alberto</creator><creator>Visentainer, Jeane Eliete Laguila</creator><general>Sociedade Brasileira de Medicina Tropical</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>3V.</scope><scope>7QO</scope><scope>7RV</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>CLZPN</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>HCIFZ</scope><scope>K9-</scope><scope>K9.</scope><scope>KB0</scope><scope>M0R</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M2P</scope><scope>MBDVC</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>20091101</creationdate><title>DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors</title><author>Cardozo, Daniela Maira ; Guelsin, Gláucia Andréia ; Clementino, Samaia Laface ; Melo, Fabiano Cavalcante de ; Braga, Marco Antônio ; Souza, Cleonice de ; Moliterno, Ricardo Alberto ; Visentainer, Jeane Eliete Laguila</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p168t-b4288d5dc2ae0c987181aaa7cbe89538dff2263949255d3f8e85accf8179aebd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>por</language><creationdate>2009</creationdate><topic>Blood</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA - 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Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes.</abstract><cop>Brazil</cop><pub>Sociedade Brasileira de Medicina Tropical</pub><pmid>20209349</pmid><doi>10.1590/S0037-86822009000600008</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals |
| subjects | Blood Deoxyribonucleic acid DNA DNA - blood DNA - isolation & purification Genotype HLA Antigens - genetics Humans Luminescent Measurements Polymerase Chain Reaction - methods Receptors, KIR - genetics |
| title | DNA extraction from coagulated human blood for application in genotyping techniques for human leukocyte antigen and immunoglobulin-like receptors |
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