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To recover this information, the linkages created must be compatible with polymerases. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Herein, a versatile ligation platform based on the formation of urea and squaramide artificial backbones from minimally modified 3\u2032\u2010 and 5\u2032\u2010amino oligonucleotides is described. One\u2010pot ligation gives a urea linkage with excellent read\u2010through speed, or a squaramide linkage that is read\u2010through under selective conditions. The squaramide linkage can be broken and reformed on demand, while stable pre\u2010activated precursor oligonucleotides expand the scope of the ligation reaction to reagent\u2010free, mild conditions. The utility of our system is demonstrated by replacing the enzymatically biased RNA\u2010to\u2010DNA reverse transcription step of RT\u2010qPCR with a rapid nucleic\u2010acid\u2010template\u2010dependent DNA chemical ligation system, that allows direct RNA detection.<\/jats:p>","DOI":"10.1002\/anie.202000209","type":"journal-article","created":{"date-parts":[[2020,3,10]],"date-time":"2020-03-10T03:21:18Z","timestamp":1583810478000},"page":"11416-11422","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Squaramides and Ureas: A Flexible Approach to Polymerase\u2010Compatible Nucleic Acid Assembly"],"prefix":"10.1002","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5206-161X","authenticated-orcid":false,"given":"Arun","family":"Shivalingam","sequence":"first","affiliation":[{"name":"Department of Chemistry University of Oxford Chemistry Research Laboratory  12 Mansfield Road Oxford OX1 3TA UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6359-4534","authenticated-orcid":false,"given":"Lapatrada","family":"Taemaitree","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Oxford Chemistry Research Laboratory  12 Mansfield Road Oxford OX1 3TA UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8706-1292","authenticated-orcid":false,"given":"Afaf H.","family":"El\u2010Sagheer","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Oxford Chemistry Research Laboratory  12 Mansfield Road Oxford OX1 3TA UK"},{"name":"Department of Science and Mathematics Suez University Faculty of Petroleum and Mining Engineering  Suez 43721 Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6538-3036","authenticated-orcid":false,"given":"Tom","family":"Brown","sequence":"additional","affiliation":[{"name":"Department of Chemistry University of Oxford Chemistry Research Laboratory  12 Mansfield Road Oxford OX1 3TA UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,5,7]]},"reference":[{"key":"e_1_2_6_1_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.211"},{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-062917-012550"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0805130105"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja808558a"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2007.104"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchem.2495"},{"key":"e_1_2_6_7_1","unstructured":"P. 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