Open AccessIn Vitro Selection Using Modified or Unnatural Nucleotides
Author(s) -
Stovall Gwendolyn M.,
Bedenbaugh Robert S.,
Singh Shruti,
Meyer Adam J.,
Hatala Paul J.,
Ellington Andrew D.,
Hall Bradley
Publication year - 2014
Publication title -
current protocols in nucleic acid chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.306
H-Index - 17
eISSN - 1934-9289
pISSN - 1934-9270
DOI - 10.1002/0471142700.nc0906s56
Subject(s) - selection (genetic algorithm) , nucleotide , in vitro , computational biology , chemistry , computer science , biology , combinatorial chemistry , genetics , artificial intelligence , gene
Incorporation of modified nucleotides into in vitro RNA or DNA selections offers many potential advantages, such as the increased stability of selected nucleic acids against nuclease degradation, improved affinities, expanded chemical functionality, and increased library diversity. 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 evaluating and optimizing transcription reactions, as well as confirming the incorporation of the modified nucleotides; protocols for evaluating modified nucleotide transcripts as template in reverse transcription reactions; protocols for the evaluation of the fidelity of modified nucleotides in the replication and the regeneration of the pool; and a protocol to compare modified nucleotide pools and selection conditions. Curr. Protoc. Nucleic Acid Chem . 56:9.6.1‐9.6.33. © 2014 by John Wiley & Sons, Inc.