| Autor: |
Carmen Grandi, Martin Emmaneel, Frank H.T. Nelissen, Maike M.K. Hansen |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
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| Zdroj: |
STAR Protocols, Vol 5, Iss 3, Pp 103284- (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2666-1667 |
| DOI: |
10.1016/j.xpro.2024.103284 |
| Popis: |
Summary: Quantifying the number of proteins that interact with mRNAs, in particular with poly(A) tails of mRNAs, is crucial for understanding gene regulation. Biochemical assays offer significant advantages for this purpose. Here, we present a protocol for synthesizing mRNAs with accurate, length-specific poly(A) tails through a PCR-based approach. We also describe steps for an in vitro (i.e., cell-free) approach for visualizing the sequential binding of Cytoplasmic Poly(A)-Binding Proteins (PABPCs) to these poly(A) tails. We detail quality control steps throughout the procedure.For complete details on the use and execution of this protocol, please refer to Grandi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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