Phosphate-Methylated Oligonucleotides as a Novel Primer for PCR and RT-PCR

Yu Hsuan Chang, Meng Wei Wu, Yi Ju Chen, Cao An Vu, Ching Ya Hong, Wen Yih Chen

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

This chapter introduces neutralized DNA (nDNA) as a novel design for the primers of PCR and RT-PCR by methylating phosphate groups of some oligonucleotides in their structures. It starts with an introduction of the nDNA which possesses an electrically chimeric neutral backbone as well as the proposed standards in designing nDNA as a novel primer for PCR and RT-PCR, concluded from various experimental results presented afterward. The primary content comprises empirical data from PCR to compare nDNA and unmodified DNA as primers in terms of ability to distinguish and amplify mismatch templates, activities of polymerase enzymes, melting temperature of double-stranded sequences, and the trials and discussions on various modified positions of the nDNA primers. In summary, nDNA exhibited outstanding performance as a primer for PCR and RT-PCR, compared to unmodified DNA, and is expected to be expanded in diverse applications which require enhanced specificity.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages261-273
Number of pages13
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2392
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • PCR
  • RT-PCR
  • nDNA

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