ArticleChembiochem : a European journal of chemical biology2020
Analysis of Modified Nucleotide Aptamer Library Generated by Thermophilic DNA Polymerases.
Article in Chembiochem : a European journal of chemical biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 10 citations in OpenAlex.
- Optimized Method for the Synthesis of Alkyne-Modified 2'-Deoxynucleoside Triphosphates.Molecules (Basel, Switzerland) · 2024Article
- MultiplexedNucleic acid therapeutics · 2024Article
- Tryptophan-like side chain holding aptamers inhibit respiratory syncytial virus infection of lung epithelial cells.Scientific reports · 2023Article
- Oligo replication advantage driven by GC content and Gibbs free energy.Biotechnology letters · 2022Article
- Enzymatic synthesis of hypermodified DNA polymers for sequence-specific display of four different hydrophobic groups.Nucleic acids research · 2020Article
- Analysis of Modified Nucleotide Aptamer Library Generated by Thermophilic DNA Polymerases.Chembiochem : a European journal of chemical biology · 2020Article
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Authors and funding
2 authors.
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Abstract
One of the pivotal steps in aptamer selection is the amplification of target-specific oligonucleotides by thermophilic DNA polymerases; it can be a challenging task if nucleic acids possessing modified nucleotides are to be amplified. Hence, the identification of compatible DNA polymerase and modified nucleotide pairs is necessary for effective selection of aptamers with unnatural nucleotides. We present an in-depth study of using 5-indolyl-AA-dUTP (TAdUTP) to generate oligonucleotide libraries for aptamer selection. We found that, among the eight studied DNA polymerases, only Vent(exo-) and KOD XL are capable of adapting TAdUTP, and that replacing dTTP did not have a significant effect on the productivity of KOD XL. We demonstrated that water-in-oil emulsion PCR is suitable for the generation of aptamer libraries of modified nucleotides. Finally, high-throughput sequence analysis showed that neither the error rate nor the PCR bias was significantly affected by using TAdUTP. In summary, we propose that KOD XL and TAdUTP could be effectively used for aptamer selection without distorting the sequence space of random oligonucleotide libraries.
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