Nicking of DNA using boronated nucleotides
AssigneeBecton, Dickinson and Company
InventorsMelinda S. Fraiser, George Terrance Walker
Life of this patent
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Abstract
.alpha.-Boronated deoxynucleoside triphosphates, when incorporated into a double-stranded restriction endonuclease recognition/cleavage site for a restriction endonuclease, induce nicking by the restriction endonuclease. .alpha.-Boronated deoxynucleoside triphosphates (dNTP.alpha.BH.sub.3) are therefore useful as nucleotide analogs in SDA to produce the nickable hemimodified restriction endonuclease recognition/cleavage site required to sustain the amplification reaction.
Claims10
A method for amplifying a target nucleic acid sequence by SDA comprising amplifying the target nucleic acid sequence in an SDA reaction wherein an alpha-boronated deoxynucleoside triphosphate is incorporated into a double-stranded recognition site for a restriction endonuclease, thereby producing a hemimodified restriction endonuclease recognition site which is nicked by the restriction endonuclease during the SDA reaction.
A method for producing nickable double-stranded DNA comprising incorporating an alpha-boronated deoxynucleoside triphosphate into a double-stranded recognition site for a restriction endonuclease to produce a hemimodified recognition site, thereby rendering the recognition site nickable by the restriction endonuclease.
Dependencies are inferred from the claim text.