Severity: Warning
Message: fopen(/var/lib/php/sessions/ci_sessionpebse0gtnppc74fno5v1pcim0564l0ng): Failed to open stream: No space left on device
Filename: drivers/Session_files_driver.php
Line Number: 177
Backtrace:
File: /var/www/html/index.php
Line: 316
Function: require_once
Severity: Warning
Message: session_start(): Failed to read session data: user (path: /var/lib/php/sessions)
Filename: Session/Session.php
Line Number: 137
Backtrace:
File: /var/www/html/index.php
Line: 316
Function: require_once
Severity: Warning
Message: file_get_contents(https://...@remsenmedia.com&api_key=81853a771c3a3a2c6b2553a65bc33b056f08&a=1): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
Filename: helpers/my_audit_helper.php
Line Number: 197
Backtrace:
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 197
Function: file_get_contents
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 271
Function: simplexml_load_file_from_url
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 1057
Function: getPubMedXML
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 3175
Function: GetPubMedArticleOutput_2016
File: /var/www/html/application/controllers/Detail.php
Line: 575
Function: pubMedSearch_Global
File: /var/www/html/application/controllers/Detail.php
Line: 489
Function: pubMedGetRelatedKeyword
File: /var/www/html/index.php
Line: 316
Function: require_once
RNA and RNA-binding proteins (RBPs) control multiple biological processes. The spatial and temporal arrangement of RNAs and RBPs underlies the delicate regulation of these processes. The strategy called CLIP-seq (cross-linking and immunoprecipitation) has been developed to capture endogenous protein-RNA interactions with UV cross-linking followed by immunoprecipitation. Despite the wide use of conventional CLIP-seq method in RBP study, the CLIP experiment is limited by the availability of the high-quality antibodies, potential contaminants from the co-purified RBPs, requirement of isotope manipulation, and potential loss of information during tedious experimental procedure. Here we described a modified CLIP-seq method called CLIP-seq using the FLAG-Biotin tag tandem purification. Through tandem purification and stringent wash condition, almost all the interacting RNA-binding proteins are removed; thus the indirect interacting RNAs mediated by these co-purified RBPs are also decreased. Our CLIP-seq method allows efficient detection of direct protein-bound RNAs without SDS-PAGE and membrane transfer procedure in an isotope-free and protein-specific antibody-free manner.
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Source |
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http://dx.doi.org/10.1007/978-1-0716-0680-3_12 | DOI Listing |
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