| Title: |
Deep sequencing of non-enzymatic RNA primer extension |
| Authors: |
Duzdevich, Daniel; Carr, Christopher E; Szostak, Jack W |
| Contributors: |
Simons Foundation; National Science Foundation; National Aeronautics and Space Administration; Howard Hughes Medical Institute |
| Source: |
Nucleic Acids Research ; volume 48, issue 12, page e70-e70 ; ISSN 0305-1048 1362-4962 |
| Publisher Information: |
Oxford University Press (OUP) |
| Publication Year: |
2020 |
| Description: |
Life emerging in an RNA world is expected to propagate RNA as hereditary information, requiring some form of primitive replication without enzymes. Non-enzymatic template-directed RNA primer extension is a model of the copying step in this posited form of replication. The sequence space accessed by primer extension dictates potential pathways to self-replication and, eventually, ribozymes. Which sequences can be accessed? What is the fidelity of the reaction? Does the recently illuminated mechanism of primer extension affect the distribution of sequences that can be copied? How do sequence features respond to experimental conditions and prebiotically relevant contexts? To help answer these and related questions, we here introduce a deep-sequencing methodology for studying RNA primer extension. We have designed and vetted special RNA constructs for this purpose, honed a protocol for sample preparation and developed custom software that analyzes sequencing data. We apply this new methodology to proof-of-concept controls, and demonstrate that it works as expected and reports on key features of the sequences accessed by primer extension. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.1093/nar/gkaa400 |
| Availability: |
https://doi.org/10.1093/nar/gkaa400; http://academic.oup.com/nar/article-pdf/48/12/e70/33468485/gkaa400.pdf |
| Rights: |
http://creativecommons.org/licenses/by-nc/4.0/ |
| Accession Number: |
edsbas.E8AE6F2F |
| Database: |
BASE |