Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

Joining oligonucleotides together (ligation) is a powerful means of retrieving information at the nanoscale. In order to recover this information, the linkages created must be compatible with polymerase enzymes. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Here, a versatile ligation platform based on the formation of urea and squaramide artificial backbones from minimally modified 3'- and 5'-amino oligonucleotides is described. One-pot ligation gives a urea linkage with excellent read-through speed, or a squaramide linkage that is read-through under selective conditions. The squaramide linkage can be broken and reformed on demand, while stable pre-activated precursor oligonucleotides expand the scope of the ligation reaction to reagent-free, mild conditions. The utility of our system is demonstrated by replacing the enzymatically biased RNA-to-DNA reverse transcription step of RT-qPCR with a rapid nucleic acid template-dependent DNA chemical ligation system, an approach that allows direct RNA detection.

Original publication

DOI

10.1002/anie.202000209

Type

Journal article

Journal

Angew Chem Int Ed Engl

Publication Date

09/03/2020

Keywords

Ligation, PCR, bio-conjugation, nucleic acids, squaramide