Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro
Summary: We present a protocol for the in vitro detection of spontaneous termination of yeast RNA polymerase II (RNAPII) transcription using bead-immobilized elongation complexes (ECs). We describe the steps for EC assembly, ligation to a long transcription template, and the in vitro elongation and...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-12-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724005343 |
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| author | Zhong Han Jesper Q. Svejstrup |
| author_facet | Zhong Han Jesper Q. Svejstrup |
| author_sort | Zhong Han |
| collection | DOAJ |
| description | Summary: We present a protocol for the in vitro detection of spontaneous termination of yeast RNA polymerase II (RNAPII) transcription using bead-immobilized elongation complexes (ECs). We describe the steps for EC assembly, ligation to a long transcription template, and the in vitro elongation and termination reactions. Our protocol has proven successful for identifying spontaneous termination in the yeast CYC1 terminator.For complete details on the use and execution of this protocol, please refer to Han et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-cc6c63c44efe41478af6e5bd48e82c44 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-cc6c63c44efe41478af6e5bd48e82c442024-11-20T05:07:59ZengElsevierSTAR Protocols2666-16672024-12-0154103369Protocol to detect spontaneous termination of yeast RNAPII transcription in vitroZhong Han0Jesper Q. Svejstrup1Department of Cellular and Molecular Medicine, Panum Institute, Blegdamsvej 3B, University of Copenhagen, 2200 Copenhagen N, Denmark; Corresponding authorDepartment of Cellular and Molecular Medicine, Panum Institute, Blegdamsvej 3B, University of Copenhagen, 2200 Copenhagen N, Denmark; Corresponding authorSummary: We present a protocol for the in vitro detection of spontaneous termination of yeast RNA polymerase II (RNAPII) transcription using bead-immobilized elongation complexes (ECs). We describe the steps for EC assembly, ligation to a long transcription template, and the in vitro elongation and termination reactions. Our protocol has proven successful for identifying spontaneous termination in the yeast CYC1 terminator.For complete details on the use and execution of this protocol, please refer to Han et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724005343molecular biologygene expressionprotein biochemistry |
| spellingShingle | Zhong Han Jesper Q. Svejstrup Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro STAR Protocols molecular biology gene expression protein biochemistry |
| title | Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro |
| title_full | Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro |
| title_fullStr | Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro |
| title_full_unstemmed | Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro |
| title_short | Protocol to detect spontaneous termination of yeast RNAPII transcription in vitro |
| title_sort | protocol to detect spontaneous termination of yeast rnapii transcription in vitro |
| topic | molecular biology gene expression protein biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724005343 |
| work_keys_str_mv | AT zhonghan protocoltodetectspontaneousterminationofyeastrnapiitranscriptioninvitro AT jesperqsvejstrup protocoltodetectspontaneousterminationofyeastrnapiitranscriptioninvitro |