Engineered templates and their use in single primer amplification

A method of amplifying nucleic acid encoding at least a portion of an antibody comprising:

• a) annealing a primer and a boundary oligonucleotide to a template that encodes at least a portion of an antibody, the primer having a first portion which anneals to the template and a second portion of predetermined sequence which does not anneal to the template;
• b) synthesizing a polynucleotide that is complementary to the portion of the template between the location at which the first portion of the primer anneals to the template and the portion of the template to which the boundary oligonucleotide anneals, the polynucleotide having the primer at a first end thereof and a second end;
• c) separating the polynucleotide synthesized in step (b) from the template;
• d) annealing a template oligonucleotide to the second end of the polynucleotide synthesized in step (b), the template oligonucleotide having a first portion that anneals to the second end of the polynucleotide and a second portion having the same predetermined sequence as the second portion of the primer;
• e) extending the polynucleotide synthesized in step (b) to provide a terminal portion thereof that is complementary to the predetermined sequence;
• f) amplifying the extended polynucleotide using a single primer having the predetermined sequence.

A method of producing an antibody library comprising:

• a) providing a diverse population of templates that encode at least a portion of an IgA antibody;
• b) contacting the diverse population of templates with at least one primer, the at least one primer having a first portion which anneals to the templates and a second portion of predetermined sequence which does not anneal to the templates;
• c) synthesizing polynucleotides that are complementary to the portion of the templates between the location at which the first portion of the primer anneals to the template and the end of the templates, the polynucleotides having the primer at a first end thereof and a second end;
• d) separating the polynucleotides synthesized in step (c) from the templates;
• e) annealing at least one template oligonucleotide to the second end of the polynucleotides synthesized in step (c), the at least one template oligonucleotide having a first portion that anneals to the second end of the polynucleotides and a second portion having the same predetermined sequence as the second portion of the primer;
• f) extending the polynucleotides synthesized in step (c) to provide a terminal portion thereof that is complementary to the predetermined sequence;
• and
• g) amplifying the extended polynucleotides using a single primer having the predetermined sequence.