Loading…
Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion
Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, “easier-seq”, to reverse transcribe and amplify the total RNAs, both with and w...
Saved in:
| Published in: | Analytical chemistry (Washington) 2016-11, Vol.88 (22), p.10795-10799 |
|---|---|
| Main Authors: | , , , |
| Format: | Article |
| Language: | English |
| Subjects: | |
| Citations: | Items that this one cites Items that cite this one |
| Online Access: | Get full text |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| cited_by | cdi_FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3 |
|---|---|
| cites | cdi_FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3 |
| container_end_page | 10799 |
| container_issue | 22 |
| container_start_page | 10795 |
| container_title | Analytical chemistry (Washington) |
| container_volume | 88 |
| creator | Fu, Yusi Chen, He Liu, Lu Huang, Yanyi |
| description | Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, “easier-seq”, to reverse transcribe and amplify the total RNAs, both with and without polyadenylate tails, from a single cell for transcriptome sequencing with high efficiency, reproducibility, and accuracy. By distributing the reverse transcribed cDNA molecules into 1.5 × 105 aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-pL reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. Meanwhile, with less chance to induce errors, this method can easily maintain the quality of single-cell sequencing. In addition, this polyadenylate-tail-independent method can be seamlessly applied to prokaryotic cell RNA sequencing. |
| doi_str_mv | 10.1021/acs.analchem.6b02581 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1864536472</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>4261040451</sourcerecordid><originalsourceid>FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3</originalsourceid><addsrcrecordid>eNqNkc1uGyEUhVHVqnHcvkFVIXWTzbiXH8_A0nKcHylKqzpdI4ZhYixmcGFmkbcPlp1E6iLyisX9zoHLh9A3AjMClPzUJs10r73Z2G5W1kDngnxAEzKnUJRC0I9oAgCsoBXAGTpPaQtACJDyMzqjVSUYF3yC1Nr1j97ipfUeP4RBe_znfoHX9t9oe5NneNjEMD5u8G0Kw8bGLhOLbudd64weXOix6_FvZ4J3g43FZQw7bwe86kaf8vQL-tRqn-zX4zlFf69WD8ub4u7X9e1ycVdoXsmhMLZquKmlqCXfb9BaruumFRJkU7ZESlkDqSkYzTRIaBi0jBtiLG-05KVlU3Rx6N3FkJ-eBtW5ZPJSurdhTIqIks9ZySt6CsooCJDkBJTNASiVIqM__kO3YYzZz57iLP97RjPFD5SJIaVoW7WLrtPxSRFQe60qa1UvWtVRa459P5aPdWeb19CLxwzAAdjH3y5-r_MZFWCwpA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1843016350</pqid></control><display><type>article</type><title>Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion</title><source>American Chemical Society:Jisc Collections:American Chemical Society Read & Publish Agreement 2022-2024 (Reading list)</source><creator>Fu, Yusi ; Chen, He ; Liu, Lu ; Huang, Yanyi</creator><creatorcontrib>Fu, Yusi ; Chen, He ; Liu, Lu ; Huang, Yanyi</creatorcontrib><description>Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, “easier-seq”, to reverse transcribe and amplify the total RNAs, both with and without polyadenylate tails, from a single cell for transcriptome sequencing with high efficiency, reproducibility, and accuracy. By distributing the reverse transcribed cDNA molecules into 1.5 × 105 aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-pL reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. Meanwhile, with less chance to induce errors, this method can easily maintain the quality of single-cell sequencing. In addition, this polyadenylate-tail-independent method can be seamlessly applied to prokaryotic cell RNA sequencing.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.6b02581</identifier><identifier>PMID: 27783484</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Amplification ; Biochemistry ; Cells ; Deoxyribonucleic acid ; Distributing ; DNA ; Droplets ; Efficiency ; Gene sequencing ; Molecules ; Reactors ; Reproducibility ; Ribonucleic acid ; Ribonucleic acids ; RNA</subject><ispartof>Analytical chemistry (Washington), 2016-11, Vol.88 (22), p.10795-10799</ispartof><rights>Copyright © 2016 American Chemical Society</rights><rights>Copyright American Chemical Society Nov 15, 2016</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3</citedby><cites>FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27783484$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fu, Yusi</creatorcontrib><creatorcontrib>Chen, He</creatorcontrib><creatorcontrib>Liu, Lu</creatorcontrib><creatorcontrib>Huang, Yanyi</creatorcontrib><title>Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, “easier-seq”, to reverse transcribe and amplify the total RNAs, both with and without polyadenylate tails, from a single cell for transcriptome sequencing with high efficiency, reproducibility, and accuracy. By distributing the reverse transcribed cDNA molecules into 1.5 × 105 aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-pL reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. Meanwhile, with less chance to induce errors, this method can easily maintain the quality of single-cell sequencing. In addition, this polyadenylate-tail-independent method can be seamlessly applied to prokaryotic cell RNA sequencing.</description><subject>Amplification</subject><subject>Biochemistry</subject><subject>Cells</subject><subject>Deoxyribonucleic acid</subject><subject>Distributing</subject><subject>DNA</subject><subject>Droplets</subject><subject>Efficiency</subject><subject>Gene sequencing</subject><subject>Molecules</subject><subject>Reactors</subject><subject>Reproducibility</subject><subject>Ribonucleic acid</subject><subject>Ribonucleic acids</subject><subject>RNA</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNqNkc1uGyEUhVHVqnHcvkFVIXWTzbiXH8_A0nKcHylKqzpdI4ZhYixmcGFmkbcPlp1E6iLyisX9zoHLh9A3AjMClPzUJs10r73Z2G5W1kDngnxAEzKnUJRC0I9oAgCsoBXAGTpPaQtACJDyMzqjVSUYF3yC1Nr1j97ipfUeP4RBe_znfoHX9t9oe5NneNjEMD5u8G0Kw8bGLhOLbudd64weXOix6_FvZ4J3g43FZQw7bwe86kaf8vQL-tRqn-zX4zlFf69WD8ub4u7X9e1ycVdoXsmhMLZquKmlqCXfb9BaruumFRJkU7ZESlkDqSkYzTRIaBi0jBtiLG-05KVlU3Rx6N3FkJ-eBtW5ZPJSurdhTIqIks9ZySt6CsooCJDkBJTNASiVIqM__kO3YYzZz57iLP97RjPFD5SJIaVoW7WLrtPxSRFQe60qa1UvWtVRa459P5aPdWeb19CLxwzAAdjH3y5-r_MZFWCwpA</recordid><startdate>20161115</startdate><enddate>20161115</enddate><creator>Fu, Yusi</creator><creator>Chen, He</creator><creator>Liu, Lu</creator><creator>Huang, Yanyi</creator><general>American Chemical Society</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20161115</creationdate><title>Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion</title><author>Fu, Yusi ; Chen, He ; Liu, Lu ; Huang, Yanyi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Amplification</topic><topic>Biochemistry</topic><topic>Cells</topic><topic>Deoxyribonucleic acid</topic><topic>Distributing</topic><topic>DNA</topic><topic>Droplets</topic><topic>Efficiency</topic><topic>Gene sequencing</topic><topic>Molecules</topic><topic>Reactors</topic><topic>Reproducibility</topic><topic>Ribonucleic acid</topic><topic>Ribonucleic acids</topic><topic>RNA</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fu, Yusi</creatorcontrib><creatorcontrib>Chen, He</creatorcontrib><creatorcontrib>Liu, Lu</creatorcontrib><creatorcontrib>Huang, Yanyi</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fu, Yusi</au><au>Chen, He</au><au>Liu, Lu</au><au>Huang, Yanyi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2016-11-15</date><risdate>2016</risdate><volume>88</volume><issue>22</issue><spage>10795</spage><epage>10799</epage><pages>10795-10799</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Prevalent single cell RNA amplification and sequencing chemistries mainly focus on polyadenylated RNAs in eukaryotic cells by using oligo(dT) primers for reverse transcription. We develop a new RNA amplification method, “easier-seq”, to reverse transcribe and amplify the total RNAs, both with and without polyadenylate tails, from a single cell for transcriptome sequencing with high efficiency, reproducibility, and accuracy. By distributing the reverse transcribed cDNA molecules into 1.5 × 105 aqueous droplets in oil, the cDNAs are isothermally amplified using random primers in each of these 65-pL reactors separately. This new method greatly improves the ease of single-cell RNA sequencing by reducing the experimental steps. Meanwhile, with less chance to induce errors, this method can easily maintain the quality of single-cell sequencing. In addition, this polyadenylate-tail-independent method can be seamlessly applied to prokaryotic cell RNA sequencing.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>27783484</pmid><doi>10.1021/acs.analchem.6b02581</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2700 |
| ispartof | Analytical chemistry (Washington), 2016-11, Vol.88 (22), p.10795-10799 |
| issn | 0003-2700 1520-6882 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1864536472 |
| source | American Chemical Society:Jisc Collections:American Chemical Society Read & Publish Agreement 2022-2024 (Reading list) |
| subjects | Amplification Biochemistry Cells Deoxyribonucleic acid Distributing DNA Droplets Efficiency Gene sequencing Molecules Reactors Reproducibility Ribonucleic acid Ribonucleic acids RNA |
| title | Single Cell Total RNA Sequencing through Isothermal Amplification in Picoliter-Droplet Emulsion |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-08T03%3A54%3A16IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Single%20Cell%20Total%20RNA%20Sequencing%20through%20Isothermal%20Amplification%20in%20Picoliter-Droplet%20Emulsion&rft.jtitle=Analytical%20chemistry%20(Washington)&rft.au=Fu,%20Yusi&rft.date=2016-11-15&rft.volume=88&rft.issue=22&rft.spage=10795&rft.epage=10799&rft.pages=10795-10799&rft.issn=0003-2700&rft.eissn=1520-6882&rft.coden=ANCHAM&rft_id=info:doi/10.1021/acs.analchem.6b02581&rft_dat=%3Cproquest_cross%3E4261040451%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-a479t-ce7d4cb98b941520fe4abdf8909d6f1999b01b20ca3a090d30f34c1ce4da946e3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1843016350&rft_id=info:pmid/27783484&rfr_iscdi=true |