Document Detail

New Pyrosequencing Method to Analyze the Function of the Klenow Fragment (EXO(-)) for Unnatural Nucleic Acids: Pyrophosphorolysis and Incorporation Efficiency.
MedLine Citation:
PMID:  22908951     Owner:  NLM     Status:  In-Data-Review    
The incorporation of deoxynucleoside triphosphates (dNTPs) catalyzed by polymerases is conventionally examined using gel electrophoresis autoradiography. Here, we studied an alternative method, pyrosequencing, to verify the incorporation of dNTPs containing unnatural nucleotides by polymerases. We found that the pyrosequencing method more rapidly and easily confirmed the incorporation of dNTPs than the conventional method, especially in the presence of low-efficiency dNTP polymerases. Furthermore, the method can detect the pyrophosphorolysis reaction just before the position of the unnatural nucleic acid, and the efficiency of incorporation just after it.
Takeshi Hanami; Rieko Oyama; Masayoshi Itoh; Ayako Yasunishi-Koyama; Yoshihide Hayashizaki
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Nucleosides, nucleotides & nucleic acids     Volume:  31     ISSN:  1532-2335     ISO Abbreviation:  Nucleosides Nucleotides Nucleic Acids     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-08-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100892832     Medline TA:  Nucleosides Nucleotides Nucleic Acids     Country:  United States    
Other Details:
Languages:  eng     Pagination:  608-15     Citation Subset:  IM    
a Omics Science Center, RIKEN Yokohama Institute , Tsurumi-ku , Yokohama , Japan.
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