Modified nucleotides

CAFC
  • US 7,771,973 B2
  • Filed: 06/01/2009
  • Issued: 08/10/2010
  • Est. Priority Date: 12/23/2002
  • Status: Active Grant
First Claim
Patent Images

1. A method for determining the sequence of a target single-stranded polynucleotide, comprising monitoring the sequential incorporation of complementary nucleotides, wherein at least one incorporation is of a nucleotide having a removable 3′

  • -OH blocking group covalently attached thereto, such that the 3′

    carbon atom has attached a group of the structure


    O—

    Zwherein Z is any of allyl, —

    C(R′

    )2

    N3, —

    C(RIV)2

    O—

    R″

    , —

    C(R′

    )2

    N(R″

    )2, —

    C(R′

    )2

    N(H)R″

    , or —

    C(RIV)2

    S—

    R″

    ,wherein —

    C(RIV)2

    O—

    R″

    is of the formula —

    CR4(R5)—

    O—

    CR4(R5)—

    OR6 or of the formula —

    CR4(R5)—

    O—

    CR4(R5)—

    SR6; and

    wherein —

    C(RIV)2

    S—

    R″

    is of the formula —

    CR4(R5)—

    S—

    CR4(R5)—

    OR6 or of the formula —

    CR4(R5)—

    S—

    CR4(R5)—

    SR6;

    each R″

    is or is part of a removable protecting group;

    each R′

    is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group;

    or (R′

    )2 represents an alkylidene group of formula ═

    C(R′



    )2 wherein each R′



    may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups;

    each R4 and R5 is independently a hydrogen atom or an alkyl group; and

    R6 is alkyl, cycloalkyl, alkenyl, cycloalkenyl or benzyl,and wherein the blocking group is removed prior to introduction of the next complementary nucleotide.

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