In vitro selection using modified or unnatural nucleotides.
The use of modified nucleotides in an RNA or DNA pool to be used for in vitro selection offers many potential advantages, such as the increased stability of the selected nucleic acid against nuclease degradation. This unit provides useful information and protocols for in vitro selection using modified nucleotides. It includes a discussion of when to use modified nucleotides; protocols for preparing a modified RNA pool and verifying its suitability for in vitro selection; and protocols for selecting and amplifying a functionally enriched pool.
Knudsen
Scott M
SM
University of Texas, Austin, Texas, USA.
Robertson
Michael P
MP
Ellington
Andrew D
AD
D002851:Chromatography, High Pressure Liquid; D004261:DNA Replication / Q000502:physiology; D019020:Directed Molecular Evolution / Q000379:methods*; D005069:Evaluation Studies as Topic; D009711:Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry* / Q000302:isolation & purification; D015202:Protein Engineering / Q000379:methods; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000502:physiology
D009711:Nucleotides; D012313:RNA; D012194:RNA-Directed DNA Polymerase