Topoisomerase linker-mediated amplification methods
AssigneesInvitrogen Corporation, South Carolina Research Institute
InventorsJohn Heyman, Gabor Szalai, Michael Felder
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Abstract
A Vaccinia topoisomerase-adapted linker, which contains an oligonucleotide primer binding site of known sequence, useful for specifically joining the linker to the end of a polynucleotide of unknown sequence and allowing subsequent PCR amplification of the polynucleotide or DNA is provided. Kits containing the invention Vaccinia topoisomerase-adapted linker and one or more linker-specific oligonucleotides for annealing to the linker in PCR are also provided. In addition, the invention provides methods for using the invention linkers in linker-mediated PCR amplification procedures for isolation and optional sequencing of isolated PCR amplification products.
Claims39
A double stranded topoisomerase-adapted linker comprising a duplex oligonucleotide linker, wherein a first oligonucleotide is annealed to a second oligonucleotide that is phosphorylated at the 5′ end thereof, and wherein the linker further comprises a compatible site-specific topoisomerase enzyme covalently attached to the first oligonucleotide at a single base 3′ T overhang.
A topoisomerase linker-specific oligonucleotide having a sequence according to SEQ ID NO:
- 11.
A topoisomerase linker-specific oligonucleotide having a sequence according to SEQ ID NO:
- 12.
Dependencies are inferred from the claim text.