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AssigneeBehringwerke Aktiengesellschaft
InventorsMaureen Laney, Yan Chen, Edwin F. Ullman, Karen M. Hahnenberger
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A method is disclosed for extending a primer to produce a single stranded polydeoxynucleotide that has two or more defined sequences. A combination is provided which comprises a template polynucleotide, a blocker polynucleotide, a primer polynucleotide and a polynucleotide Q. The template polynucleotide has three sequences T1, T2 and T3 wherein T1 is non-contiguous and 3' of T3 and wherein the 5' end of T3 is 5' of the 5' end of T2. The primer polynucleotide has a second defined sequence at its 3' end that is hybridizable with T1. The blocker polynucleotide has sequence B1 that is hybridizable with T3. Polynucleotide Q has sequences S1 and S2 wherein S1 is 3' of S2 and homologous with T2 and S2 is complementary to a first defined sequence that is to be introduced at the 3' end of the polynucleotide primer, when it is extended during the method of the invention. Polynucleotide Q is either attached to the 5' end of the blocker polynucleotide or present as a separate reagent. The primer is extended along the template polynucleotide and along at least a portion of sequence T2 and thereafter along the polynucleotide Q to give a single stranded polynucleotide having two or more defined sequences.
A method for producing from a polynucleotide primer a single stranded polydeoxynucleotide having two different defined sequences P1 and P'2, said method comprising the steps of:
An isolated polynucleotide primer comprising a single strand of DNA comprised of a sequence T3 and a sequence T2 wherein the 3' end of said T3 is contiguous with the 5' end of T2, said single strand being complexed to a blocker DNA sequence comprised of a sequence B1 complementary to said T3 and a sequence S1 which is 5' of B1 and substantially identical to said T2.
A kit comprising in packaged combination:
Dependencies are inferred from the claim text.