Extraction of high-quality RNA from human articular cartilage
Extracting high-quality RNA from articular cartilage is challenging due to low cellularity and high proteoglycan content. This problem hinders efficient application of RNA sequencing (RNA-seq) analysis in studying cartilage homeostasis. Here we developed a method that purifies high-quality RNA direc...
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| Veröffentlicht in: | Analytical biochemistry 2017-02, Vol.518, p.134-138 |
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| container_title | Analytical biochemistry |
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| creator | Le Bleu, Heather K. Kamal, Fadia A. Kelly, Meghan Ketz, John P. Zuscik, Michael J. Elbarbary, Reyad A. |
| description | Extracting high-quality RNA from articular cartilage is challenging due to low cellularity and high proteoglycan content. This problem hinders efficient application of RNA sequencing (RNA-seq) analysis in studying cartilage homeostasis. Here we developed a method that purifies high-quality RNA directly from cartilage. Our method optimized the collection and homogenization steps so as to minimize RNA degradation, and modified the conventional TRIzol protocol to enhance RNA purity. Cartilage RNA purified using our method has appropriate quality for RNA-seq experiments including an RNA integrity number of ∼8. Our method also proved efficient in extracting high-quality RNA from subchondral bone. |
| doi_str_mv | 10.1016/j.ab.2016.11.018 |
| format | Article |
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This problem hinders efficient application of RNA sequencing (RNA-seq) analysis in studying cartilage homeostasis. Here we developed a method that purifies high-quality RNA directly from cartilage. Our method optimized the collection and homogenization steps so as to minimize RNA degradation, and modified the conventional TRIzol protocol to enhance RNA purity. Cartilage RNA purified using our method has appropriate quality for RNA-seq experiments including an RNA integrity number of ∼8. Our method also proved efficient in extracting high-quality RNA from subchondral bone.</description><subject>Cartilage, Articular - chemistry</subject><subject>Humans</subject><subject>Knee Joint - chemistry</subject><subject>Male</subject><subject>RNA - chemistry</subject><subject>RNA - isolation & purification</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM1r3DAQxUVp6G7S3nsqPvZiR-MPSS60EEKaBEICZe9iJI93tdjWRrJD8t_Xy25Dc8hpBua9N48fY1-BZ8BBnG8zNFk-bxlAxkF9YEvgtUh5weuPbMk5L9Jc1HLBTmPccg5QVuITW-SyhgJEuWQ_r57HgHZ0fkh8m2zcepM-Tti58SX5c3-RtMH3yWbqcUgwjM5OHYbE7tcO1_SZnbTYRfpynGds9ftqdXmT3j1c315e3KW2LOWYCiPINNZCI0RTGoWVshI45aY0tZFWKhSVVYBYqMZyhXUrJRlqAaocm-KM_TrE7ibTU2NpmDt3ehdcj-FFe3T67WVwG732T7rKoZKVnAO-HwOCf5wojrp30VLX4UB-ihpUWalCyYLPUn6Q2uBjDNS-vgGu99D1VqPRe-gaQM_QZ8u3_-u9Gv5RngU_DgKaGT05CjpaR4OlxgWyo268ez_9L58kkus</recordid><startdate>20170201</startdate><enddate>20170201</enddate><creator>Le Bleu, Heather K.</creator><creator>Kamal, Fadia A.</creator><creator>Kelly, Meghan</creator><creator>Ketz, John P.</creator><creator>Zuscik, Michael J.</creator><creator>Elbarbary, Reyad A.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20170201</creationdate><title>Extraction of high-quality RNA from human articular cartilage</title><author>Le Bleu, Heather K. ; Kamal, Fadia A. ; Kelly, Meghan ; Ketz, John P. ; Zuscik, Michael J. ; Elbarbary, Reyad A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c447t-6b6ebdcc1d66d4b8a58c710e2b4b9b7c78a65c81aa38dc08a9f77ebef1152ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Cartilage, Articular - chemistry</topic><topic>Humans</topic><topic>Knee Joint - chemistry</topic><topic>Male</topic><topic>RNA - chemistry</topic><topic>RNA - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Le Bleu, Heather K.</creatorcontrib><creatorcontrib>Kamal, Fadia A.</creatorcontrib><creatorcontrib>Kelly, Meghan</creatorcontrib><creatorcontrib>Ketz, John P.</creatorcontrib><creatorcontrib>Zuscik, Michael J.</creatorcontrib><creatorcontrib>Elbarbary, Reyad A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Le Bleu, Heather K.</au><au>Kamal, Fadia A.</au><au>Kelly, Meghan</au><au>Ketz, John P.</au><au>Zuscik, Michael J.</au><au>Elbarbary, Reyad A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Extraction of high-quality RNA from human articular cartilage</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>2017-02-01</date><risdate>2017</risdate><volume>518</volume><spage>134</spage><epage>138</epage><pages>134-138</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>Extracting high-quality RNA from articular cartilage is challenging due to low cellularity and high proteoglycan content. 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| ispartof | Analytical biochemistry, 2017-02, Vol.518, p.134-138 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Cartilage, Articular - chemistry Humans Knee Joint - chemistry Male RNA - chemistry RNA - isolation & purification |
| title | Extraction of high-quality RNA from human articular cartilage |
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