Methods and kits to enrich for desired nucleic acid sequences

A method for producing a vector comprising a desired nucleic acid insert from a sample comprising a plurality of vectors, the method comprising:

• generating first and second extension products from first and second primers annealed to a vector in the sample, wherein the vector comprises a desired nucleic acid insert, wherein the first and second primers anneal to complementary strands of a vector comprising the desired nucleic acid insert, wherein one or both of the first or second primers contain a nucleic acid sequence found in the desired nucleic acid insert or its complement;
• providing a nucleic acid comprising an additional vector sequence, wherein the nucleic acids of the first and second extension products together with the additional vector sequence comprise a sequence for replication in a host and comprise the desired nucleic acid insert, and wherein the first and second extension products are capable of annealing to the additional vector sequence;
• optionally repeating the step of generating first and second extension products;
• and
• combining and annealing the first and second extension products and the nucleic acid comprising an additional vector sequence to form a partially double-stranded, replication-competent vector.

A method for producing a vector from a sample comprising a library of nucleic acid sequences, comprising:

• denaturing the nucleic acids in the sample;
• performing one or more amplification reactions by adding amplification reagents and primers such that a first primer pair is capable of initiating the amplification of a first extension product made with a nucleic acid comprising a vector, and a second primer pair is capable of initiating the amplification of a second extension product made with the nucleic acid comprising the vector, whereby the first and second extension products together comprise a sequence for replication in a host and a desired nucleic acid insert, and wherein the first and second extension products comprise nucleic acid regions capable of allowing the first and second extension products to anneal to each other at both their terminal ends;
• and
• annealing the first and second extension products to form a replication-competent vector.

A method for enriching the amount of a desired nucleic acid in a sample comprising a mixture of nucleic acids comprising:

• producing a vector comprising the desired nucleic acid;
• generating first and second extension products from first and second primers annealed to the vector in the sample, wherein the vector comprises the desired nucleic acid, wherein the first and second primers anneal to complementary strands of a vector comprising the desired nucleic acid insert, wherein one or both of the first or second primers comprise a nucleic acid sequence found in the desired nucleic acid insert or its complement, wherein the first and second extension products are capable of annealing to each other at both of their terminal ends, and wherein the nucleic acids of the first and second extension products together comprise a sequence for replication in a host and the desired nucleic acid insert;
• optionally repeating the generating first and second extension products;
• and
• combining and annealing the first and second extension products to form a partially double-stranded, replication-competent vector.

A method for enriching the amount of a desired nucleic acid in a sample comprising a mixture of nucleic acids, comprising:

• producing a vector comprising the desired nucleic acid;
• generating first and second extension products from first and second primers annealed to a vector in the sample, wherein the vector comprises a desired nucleic acid inser, wherein the first and second primers anneal to complementary strands of the vector comprising a desired nucleic acid insert, wherein one or both of the first or second primers contain a nucleic acid sequence found in the desired nucleic acid insert or its complement;
• providing a nucleic acid comprising an additional vector sequence, wherein the nucleic acids of the first and second extension products together with the additional vector sequence comprise a sequence for replication in a host and comprise the desired nucleic acid insert, and wherein the first and second extension products are capable of annealing to the additional vector sequence;
• optionally repeating the step of generating first and second extension products;
• and
• combining and annealing the first and second extension products and the nucleic acid comprising an additional vector sequence to form a partially stranded—stranded, replication-competent vector.

A method for enriching the amount of a desired nucleic acid in a sample comprising a library of nucleic acid sequences, comprising:

• denaturing the nucleic acids in the sample;
• performing one or more amplification reactions by adding amplification reagents and primers such that a first primer pair is capable of initiating the amplification of a first extension product made with a vector comprising a desired nucleic acid, and a second primer pair is capable of initiating the amplification of a second extension product made with the vector comprising the desired nucleic acid, whereby the first and second extension products together comprise a sequence for replication in a host and a desired nucleic acid, and wherein the first and second extension products comprise nucleic acid regions capable of allowing the first and second extension products to anneal to each other;
• and
• annealing the first and second extension products to form a replication-competent vector.