Inosine 5'-triphosphate can dramatically increase the yield of NASBA products targeting GC-rich and intramolecular base-paired viroid RNA | Zendy

Kenji Nakahara | Zendy; Tatsuji Hataya | Zendy; Ichiro Uyeda | Zendy

Open Access

Inosine 5'-triphosphate can dramatically increase the yield of NASBA products targeting GC-rich and intramolecular base-paired viroid RNA

Author(s) -

Kenji Nakahara,

Tatsuji Hataya,

Ichiro Uyeda

Publication year - 1998

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/26.7.1854

Subject(s) - biology , nasba , nucleic acid , base pair , inosine , rna , nucleotide , microbiology and biotechnology , virology , nucleic acid sequence , biochemistry , dna , adenosine , gene

Nucleic acid sequence-based amplification (NASBA) according to the standard protocol failed to amplify cRNA of viroids, probably because of their GC-rich and intramolecular base-paired structure. However, NASBA in the presence of inosine 5'-triphosphate successfully amplified the cRNAs to viroids in total nucleic acid extracts from citrus plants. As sequence specificity of the cRNA to viroids was confirmed by northern analysis, the amplification and fidelity of cRNAs are sufficient for the sensitive and specific detection of viroids.

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