Cozens, C; Pinheiro, VB; (2018) XNA Synthesis and Reverse Transcription by Engineered Thermophilic Polymerases. Current Protocols in Chemical Biology , 10 (3) , Article e47. 10.1002/cpch.47.

Text Cozens_XNA Synthesis and Reverse Transcription by Engineered Thermophilic Polymerases_AAM.pdf - Accepted Version Access restricted to UCL open access staff Download (504kB)

Abstract

The B‐family polymerases of hyperthermophilic archaea have proven an exceptional platform for engineering polymerases with extended substrate spectra, despite multiple mechanisms for detecting and avoiding incorporation of non‐cognate substrates. These polymerases can efficiently synthesize and reverse‐transcribe a number of xenonucleic acids (XNAs) that differ significantly from the canonical B‐form of DNA. We present here a protocol for hexitol nucleic acid (HNA) synthesis by an engineered Thermococcus gorgonarius polymerase variant, including adaptation for large‐scale synthesis and purification, and for other XNAs. We describe XNA purification and reverse transcription (with a previously reported XNA RT also based on Thermococcus gorgonarius), as well as key considerations for the characterization and optimization of XNA reactions.

Download activity - last month

Export as CSVJSONXML

Download activity - last 12 months

Export as JSONXMLCSV

Downloads by country - last 12 months

Export as CSVJSONXML

Archive Staff Only

View Item