Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles
The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only...
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| Veröffentlicht in: | Science China. Chemistry 2015-11, Vol.58 (11), p.1774-1778 |
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| creator | Wang, Jiuhai Ali, Zeeshan Wang, Nianyue Liang, Wenbiao Liu, Hongna Li, Fu Yang, Haowen He, Lei Nie, Libo He, Nongyue Li, Zhiyang |
| description | The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles （MNPs） was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease （DNase） and Ribonucleases （RNase） free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction （PCR） and reverse transcription PCR （RT-PCR） to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria. |
| doi_str_mv | 10.1007/s11426-015-5483-x |
| format | Article |
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Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles （MNPs） was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease （DNase） and Ribonucleases （RNase） free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction （PCR） and reverse transcription PCR （RT-PCR） to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.</description><identifier>ISSN: 1674-7291</identifier><identifier>EISSN: 1869-1870</identifier><identifier>DOI: 10.1007/s11426-015-5483-x</identifier><language>eng</language><publisher>Beijing: Science China Press</publisher><subject>Acids ; Bacteria ; Buffers ; Carbohydrates ; Chemistry ; Chemistry and Materials Science ; Chemistry/Food Science ; Cloning ; Deoxyribonucleic acid ; DNA ; E coli ; Molecular biology ; Nanoparticles ; Nucleic acids ; Polymerase chain reaction ; Ribonucleic acid ; RNA ; RNA提取方法 ; RT-PCR ; 二氧化硅 ; 包覆 ; 大肠杆菌 ; 磁性纳米粒子 ; 细菌DNA ; 脱氧核糖核酸酶</subject><ispartof>Science China. Chemistry, 2015-11, Vol.58 (11), p.1774-1778</ispartof><rights>Science China Press and Springer-Verlag Berlin Heidelberg 2015</rights><rights>Science China Press and Springer-Verlag Berlin Heidelberg 2015.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c413t-c78f155b6aed24570d9469975b4b7d2b7755247dba71739cd69d76a4d50b568f3</citedby><cites>FETCH-LOGICAL-c413t-c78f155b6aed24570d9469975b4b7d2b7755247dba71739cd69d76a4d50b568f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/60113X/60113X.jpg</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11426-015-5483-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2918595913?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,21388,21389,21390,21391,23256,27924,27925,33530,33703,33744,34005,34314,41488,42557,43659,43787,43805,43953,44067,51319,64385,64389,72469</link.rule.ids></links><search><creatorcontrib>Wang, Jiuhai</creatorcontrib><creatorcontrib>Ali, Zeeshan</creatorcontrib><creatorcontrib>Wang, Nianyue</creatorcontrib><creatorcontrib>Liang, Wenbiao</creatorcontrib><creatorcontrib>Liu, Hongna</creatorcontrib><creatorcontrib>Li, Fu</creatorcontrib><creatorcontrib>Yang, Haowen</creatorcontrib><creatorcontrib>He, Lei</creatorcontrib><creatorcontrib>Nie, Libo</creatorcontrib><creatorcontrib>He, Nongyue</creatorcontrib><creatorcontrib>Li, Zhiyang</creatorcontrib><title>Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles</title><title>Science China. Chemistry</title><addtitle>Sci. China Chem</addtitle><addtitle>SCIENCE CHINA Chemistry</addtitle><description>The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles （MNPs） was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease （DNase） and Ribonucleases （RNase） free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction （PCR） and reverse transcription PCR （RT-PCR） to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.</description><subject>Acids</subject><subject>Bacteria</subject><subject>Buffers</subject><subject>Carbohydrates</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chemistry/Food Science</subject><subject>Cloning</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>E coli</subject><subject>Molecular biology</subject><subject>Nanoparticles</subject><subject>Nucleic acids</subject><subject>Polymerase chain reaction</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA提取方法</subject><subject>RT-PCR</subject><subject>二氧化硅</subject><subject>包覆</subject><subject>大肠杆菌</subject><subject>磁性纳米粒子</subject><subject>细菌DNA</subject><subject>脱氧核糖核酸酶</subject><issn>1674-7291</issn><issn>1869-1870</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kMtKAzEUhgdRsGgfwF3QdTSZ3CbLeheKgpd1yCSZNmWatMkU6tub0qI7szmH8H_nh6-qLjC6xgiJm4wxrTlEmEFGGwK3R9UIN1xC3Ah0XHYuKBS1xKfVOOcFKo8QVAs2qlYffrnpBx1c3GTgtkPSZvAxgNiB-9cJ0MGC9zK7FJfgIZu5S97MvQYm9h7cTkGNQauzs6Aw2ffeaGiiHsrHUs-CG7wBQYe40qmsvcvn1Umn--zGh3lWfT0-fN49w-nb08vdZAoNxWSARjQdZqzl2tmaMoGspFxKwVraClu3QjBWU2FbLbAg0lgureCaWoZaxpuOnFVX-7urFNcblwe1iJsUSqUqIhommcSkpPA-ZVLMOblOrZJf6vStMFI7t2rvVhW3audWbQtT75lcsmHm0t_l_6DLQ9E8htm6cL9NnHPCRNMg8gN1CocM</recordid><startdate>20151101</startdate><enddate>20151101</enddate><creator>Wang, Jiuhai</creator><creator>Ali, Zeeshan</creator><creator>Wang, Nianyue</creator><creator>Liang, Wenbiao</creator><creator>Liu, Hongna</creator><creator>Li, Fu</creator><creator>Yang, Haowen</creator><creator>He, Lei</creator><creator>Nie, Libo</creator><creator>He, Nongyue</creator><creator>Li, Zhiyang</creator><general>Science China Press</general><general>Springer Nature B.V</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>~WA</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7XB</scope><scope>88I</scope><scope>8FE</scope><scope>8FG</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>KB.</scope><scope>M2P</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope></search><sort><creationdate>20151101</creationdate><title>Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles</title><author>Wang, Jiuhai ; 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Chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Jiuhai</au><au>Ali, Zeeshan</au><au>Wang, Nianyue</au><au>Liang, Wenbiao</au><au>Liu, Hongna</au><au>Li, Fu</au><au>Yang, Haowen</au><au>He, Lei</au><au>Nie, Libo</au><au>He, Nongyue</au><au>Li, Zhiyang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles</atitle><jtitle>Science China. Chemistry</jtitle><stitle>Sci. China Chem</stitle><addtitle>SCIENCE CHINA Chemistry</addtitle><date>2015-11-01</date><risdate>2015</risdate><volume>58</volume><issue>11</issue><spage>1774</spage><epage>1778</epage><pages>1774-1778</pages><issn>1674-7291</issn><eissn>1869-1870</eissn><abstract>The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles （MNPs） was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease （DNase） and Ribonucleases （RNase） free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction （PCR） and reverse transcription PCR （RT-PCR） to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.</abstract><cop>Beijing</cop><pub>Science China Press</pub><doi>10.1007/s11426-015-5483-x</doi><tpages>5</tpages></addata></record> |
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| source | ProQuest Central Essentials; ProQuest Central (Alumni Edition); ProQuest Central Student; SpringerNature Journals; ProQuest Central Korea; ProQuest Central UK/Ireland; Alma/SFX Local Collection; ProQuest Central |
| subjects | Acids Bacteria Buffers Carbohydrates Chemistry Chemistry and Materials Science Chemistry/Food Science Cloning Deoxyribonucleic acid DNA E coli Molecular biology Nanoparticles Nucleic acids Polymerase chain reaction Ribonucleic acid RNA RNA提取方法 RT-PCR 二氧化硅 包覆 大肠杆菌 磁性纳米粒子 细菌DNA 脱氧核糖核酸酶 |
| title | Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles |
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