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AssigneeLucigen Corporation
InventorsDavid A. Mead, Ronald Godiska, Thomas W. Schoenfeld, Spencer Hermanson
Estimating the term…
Vector preparations and cloning constructs suitable for use in cloning are provided. Vector preparations are double-stranded DNA molecules having two 3′ termini, each terminus having a single base pair overhang that is capable of hybridizing to a single base pair overhang on a double stranded polynucleotide sequence to be cloned. The overhang of the vector preparation is suitably a dCMP and the overhang of the polynucleotide sequence to be cloned is suitably a dGMP. In other embodiments, the overhang of the polynucleotide sequence to be cloned is any ddNTP and the corresponding overhang of the vector preparation is any base that pairs to the ddNTP. The latter embodiment is particularly suited to preparing recombinant molecules having only a single insert. Methods of cloning, methods of preparing libraries of recombinant molecules and kits for carrying out the methods are also provided.
A vector preparation comprising a double-stranded DNA molecule having two 3′ termini, each 3′ terminus comprising an overhang, each overhang comprising a single, unpaired dCMP or nucleotide analog capable of hybridizing to an unpaired dGMP.
A kit comprising instructions for use of the kit, and:
A method of preparing a double-stranded polynucleotide for cloning comprising treating the polynucleotide to provide an overhang at each 3′ terminus, each overhang comprising a single unpaired dGMP or nucleotide analog capable of hybridizing to an unpaired dCMP.
A method of preparing a double-stranded polynucleotide for cloning comprising treating the polynucleotide to provide an overhang at each 3′ terminus, each overhang comprising a single unpaired ddNMP.
Dependencies are inferred from the claim text.