Inhibition of restriction enzyme cleavage of DNA modified with 7-deaza-dGTP

DNA templates containing G+C rich regions or stable secondary structures can potentially pose problems for polymerase chain reaction amplification. Incorporation of the structure destabilizing analogue, 7-deaza-dGTP into the polymerase chain reaction reaction allows successful amplification of such...

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Bibliographic Details
Published in:Nucleic acids research 1991-05, Vol.19 (10), p.2791-2791
Main Authors: GRIME, S. K, MARTIN, R. L, HOLAWAY, B. L
Format: Article
Language:English
Subjects:
7-deaza-dGTP
Analytical, structural and metabolic biochemistry
Base Sequence
Biological and medical sciences
Deoxyguanine Nucleotides - pharmacology
DNA - metabolism
DNA Restriction Enzymes - antagonists & inhibitors
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Hydrolases
Molecular Sequence Data
polymerase chain reaction
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Summary:DNA templates containing G+C rich regions or stable secondary structures can potentially pose problems for polymerase chain reaction amplification. Incorporation of the structure destabilizing analogue, 7-deaza-dGTP into the polymerase chain reaction reaction allows successful amplification of such template sequences. It has been reported that when 7-deaza-dGTP is substituted for dGTP in a DNA-fragment, the cleavage rate by the restriction enzyme EcoRI is reduced. We find that EcoRI as well as other commonly used restriction enzymes will not cut DNA containing 7-deaza-dGTP when it is incorporated into polymerase chain reaction product.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/19.10.2791