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A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification
RNA isolation from bacteria is technically difficult due to the RNA characteristic of labile and vulnerable degradation. Many reagents were explored for cellular lysis and complete inhibition of RNase. However, the available methods for RNA isolation are either of low efficiency or time-consuming. H...
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| Published in: | Analytical and bioanalytical chemistry 2021-11, Vol.413 (26), p.6469-6477 |
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| cites | cdi_FETCH-LOGICAL-c414t-b0d88f872267ba08f1602c110eae5c17cef253bdd883467ebb18dfbf0e459d403 |
| container_end_page | 6477 |
| container_issue | 26 |
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| container_title | Analytical and bioanalytical chemistry |
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| creator | Zhao, Xiaoli Li, Yong Duan, Yake Amin, Amr Xie, Yingqiu Shi, Chao Ma, Cuiping |
| description | RNA isolation from bacteria is technically difficult due to the RNA characteristic of labile and vulnerable degradation. Many reagents were explored for cellular lysis and complete inhibition of RNase. However, the available methods for RNA isolation are either of low efficiency or time-consuming. Here, we developed a rapid and accessible protocol for RNA isolation that combined a simplified cell lysis and RNA release by formamide-based solution and RNA purification by chitosan-modified silica membrane for the first time. With this method, we obtained about ~ 28 μg of total RNA from 10
8
Escherichia coli
cells. The entire procedure can be done within 15 min without redundant pipetting steps. The purity of extracted RNA was comparable to that of commercial kits, but the cost was much lower. Furthermore, the yielded RNA was successfully used in downstream enzymatic reactions, such as reverse transcription and quantitative real-time PCR. This new method would be of benefit for an extensive range of gene expression analyses in bacterial organisms.
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| doi_str_mv | 10.1007/s00216-021-03644-6 |
| format | article |
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8
Escherichia coli
cells. The entire procedure can be done within 15 min without redundant pipetting steps. The purity of extracted RNA was comparable to that of commercial kits, but the cost was much lower. Furthermore, the yielded RNA was successfully used in downstream enzymatic reactions, such as reverse transcription and quantitative real-time PCR. This new method would be of benefit for an extensive range of gene expression analyses in bacterial organisms.
Graphical abstract</description><identifier>ISSN: 1618-2642</identifier><identifier>EISSN: 1618-2650</identifier><identifier>DOI: 10.1007/s00216-021-03644-6</identifier><identifier>PMID: 34505946</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Amides ; Analytical Chemistry ; Bacteria ; Biochemistry ; Biodegradation ; Characterization and Evaluation of Materials ; Chemical properties ; Chemistry ; Chemistry and Materials Science ; Chitin ; Chitosan ; Chitosan - analogs & derivatives ; Escherichia coli - chemistry ; Escherichia coli - genetics ; Extraction (Chemistry) ; Food Science ; Formamides - chemistry ; Gene expression ; Genetic aspects ; Laboratory Medicine ; Lysis ; Membranes, Artificial ; Methods ; Monitoring/Environmental Analysis ; Nucleic acids ; Paper in Forefront ; Purification ; Reagents ; Reverse transcription ; Ribonuclease ; Ribonucleic acid ; RNA ; RNA, Bacterial - genetics ; RNA, Bacterial - isolation & purification ; Separation ; Silica ; Silicon dioxide ; Silicon Dioxide - chemistry</subject><ispartof>Analytical and bioanalytical chemistry, 2021-11, Vol.413 (26), p.6469-6477</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2021</rights><rights>2021. Springer-Verlag GmbH Germany, part of Springer Nature.</rights><rights>COPYRIGHT 2021 Springer</rights><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2021.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c414t-b0d88f872267ba08f1602c110eae5c17cef253bdd883467ebb18dfbf0e459d403</citedby><cites>FETCH-LOGICAL-c414t-b0d88f872267ba08f1602c110eae5c17cef253bdd883467ebb18dfbf0e459d403</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34505946$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Xiaoli</creatorcontrib><creatorcontrib>Li, Yong</creatorcontrib><creatorcontrib>Duan, Yake</creatorcontrib><creatorcontrib>Amin, Amr</creatorcontrib><creatorcontrib>Xie, Yingqiu</creatorcontrib><creatorcontrib>Shi, Chao</creatorcontrib><creatorcontrib>Ma, Cuiping</creatorcontrib><title>A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification</title><title>Analytical and bioanalytical chemistry</title><addtitle>Anal Bioanal Chem</addtitle><addtitle>Anal Bioanal Chem</addtitle><description>RNA isolation from bacteria is technically difficult due to the RNA characteristic of labile and vulnerable degradation. Many reagents were explored for cellular lysis and complete inhibition of RNase. However, the available methods for RNA isolation are either of low efficiency or time-consuming. Here, we developed a rapid and accessible protocol for RNA isolation that combined a simplified cell lysis and RNA release by formamide-based solution and RNA purification by chitosan-modified silica membrane for the first time. With this method, we obtained about ~ 28 μg of total RNA from 10
8
Escherichia coli
cells. The entire procedure can be done within 15 min without redundant pipetting steps. The purity of extracted RNA was comparable to that of commercial kits, but the cost was much lower. Furthermore, the yielded RNA was successfully used in downstream enzymatic reactions, such as reverse transcription and quantitative real-time PCR. This new method would be of benefit for an extensive range of gene expression analyses in bacterial organisms.
Graphical abstract</description><subject>Amides</subject><subject>Analytical Chemistry</subject><subject>Bacteria</subject><subject>Biochemistry</subject><subject>Biodegradation</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemical properties</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chitin</subject><subject>Chitosan</subject><subject>Chitosan - analogs & derivatives</subject><subject>Escherichia coli - chemistry</subject><subject>Escherichia coli - genetics</subject><subject>Extraction (Chemistry)</subject><subject>Food Science</subject><subject>Formamides - chemistry</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Laboratory Medicine</subject><subject>Lysis</subject><subject>Membranes, Artificial</subject><subject>Methods</subject><subject>Monitoring/Environmental Analysis</subject><subject>Nucleic acids</subject><subject>Paper in Forefront</subject><subject>Purification</subject><subject>Reagents</subject><subject>Reverse transcription</subject><subject>Ribonuclease</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA, Bacterial - genetics</subject><subject>RNA, Bacterial - isolation & purification</subject><subject>Separation</subject><subject>Silica</subject><subject>Silicon dioxide</subject><subject>Silicon Dioxide - 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simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification</title><author>Zhao, Xiaoli ; Li, Yong ; Duan, Yake ; Amin, Amr ; Xie, Yingqiu ; Shi, Chao ; Ma, Cuiping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c414t-b0d88f872267ba08f1602c110eae5c17cef253bdd883467ebb18dfbf0e459d403</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Amides</topic><topic>Analytical Chemistry</topic><topic>Bacteria</topic><topic>Biochemistry</topic><topic>Biodegradation</topic><topic>Characterization and Evaluation of Materials</topic><topic>Chemical properties</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Chitin</topic><topic>Chitosan</topic><topic>Chitosan - analogs & derivatives</topic><topic>Escherichia coli - chemistry</topic><topic>Escherichia coli - 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Academic</collection><jtitle>Analytical and bioanalytical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Xiaoli</au><au>Li, Yong</au><au>Duan, Yake</au><au>Amin, Amr</au><au>Xie, Yingqiu</au><au>Shi, Chao</au><au>Ma, Cuiping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification</atitle><jtitle>Analytical and bioanalytical chemistry</jtitle><stitle>Anal Bioanal Chem</stitle><addtitle>Anal Bioanal Chem</addtitle><date>2021-11-01</date><risdate>2021</risdate><volume>413</volume><issue>26</issue><spage>6469</spage><epage>6477</epage><pages>6469-6477</pages><issn>1618-2642</issn><eissn>1618-2650</eissn><abstract>RNA isolation from bacteria is technically difficult due to the RNA characteristic of labile and vulnerable degradation. Many reagents were explored for cellular lysis and complete inhibition of RNase. However, the available methods for RNA isolation are either of low efficiency or time-consuming. Here, we developed a rapid and accessible protocol for RNA isolation that combined a simplified cell lysis and RNA release by formamide-based solution and RNA purification by chitosan-modified silica membrane for the first time. With this method, we obtained about ~ 28 μg of total RNA from 10
8
Escherichia coli
cells. The entire procedure can be done within 15 min without redundant pipetting steps. The purity of extracted RNA was comparable to that of commercial kits, but the cost was much lower. Furthermore, the yielded RNA was successfully used in downstream enzymatic reactions, such as reverse transcription and quantitative real-time PCR. This new method would be of benefit for an extensive range of gene expression analyses in bacterial organisms.
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| subjects | Amides Analytical Chemistry Bacteria Biochemistry Biodegradation Characterization and Evaluation of Materials Chemical properties Chemistry Chemistry and Materials Science Chitin Chitosan Chitosan - analogs & derivatives Escherichia coli - chemistry Escherichia coli - genetics Extraction (Chemistry) Food Science Formamides - chemistry Gene expression Genetic aspects Laboratory Medicine Lysis Membranes, Artificial Methods Monitoring/Environmental Analysis Nucleic acids Paper in Forefront Purification Reagents Reverse transcription Ribonuclease Ribonucleic acid RNA RNA, Bacterial - genetics RNA, Bacterial - isolation & purification Separation Silica Silicon dioxide Silicon Dioxide - chemistry |
| title | A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification |
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