منابع مشابه
RNA immunoprecipitation for determining RNA-protein associations in vivo.
Similar to chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP) can be used to detect the association of individual proteins with specific nucleic acid regions, in this case on RNA. Live cells are treated with formaldehyde to generate protein-RNA cross-links between molecules that are in close proximity in vivo. RNA sequences that cross-link with a given protein are isolated by i...
متن کاملMapping of psoralen cross-linked nucleotides in RNA.
A method is described for using the cross-linking reagent 4'-(hydroxy-methyl)-4,5',8-trimethylpsoralen (HMT) to map base paired regions and higher-order structure within RNA molecules. Applying this method to yeast tRNAPhe, we have specifically identified cross-links within the acceptor stem between U6 X U68, in the D-stem between C11 X C25, and in the T psi-stem between U50 X C63 and U52 X C63...
متن کاملCombination of cross-species RNA solution hybridization and immunoprecipitation aids in the cloning of RT-PCR products.
cDNA cloning by means of reverse transcription-polymerase chain reaction (RT-PCR) is a widely used technique. Ever since the first published method by Veres et al. (6), it has supplanted conventional methods of cDNA cloning due to its ease and speed. However, there are certain constraints that are associated with this technique; e.g., it requires the knowledge of a few nucleic acid sequences of...
متن کاملCross-Linking Immunoprecipitation and qPCR (CLIP-qPCR) Analysis to Map Interactions Between Long Noncoding RNAs and RNA-Binding Proteins.
Mammalian cells express a wide range of transcripts, some protein-coding RNAs (mRNA) and many noncoding (nc) RNAs. Long (l)ncRNAscan modulates protein expression patterns by regulating gene transcription, pre-mRNA splicing, mRNA export, mRNA degradation, protein translation, and protein ubiquitination. Given the growing recognition that lncRNAs have a robust impact upon gene expression, there i...
متن کاملCross-linked informofers.
The proteins of 30S RNP particles containing pre-mRNA (hnRNA) were cross-linked with bifunctional reagents (dimethyl-suberimidate and dimethyl-3,3'-dithiobispropionimidate). Further treatment with 1 or 2 M NaCl dissociates all RNA from protein. However, a significant part of protein particles--informofers being cross-linked survived high salt treatment. Their sedimentation coefficients were clo...
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ژورنال
عنوان ژورنال: BIO-PROTOCOL
سال: 2013
ISSN: 2331-8325
DOI: 10.21769/bioprotoc.398