Synthesis of Pools of Probes by Primer Extension

摘要

Provided herein is a method for making a pool of probes by primer extension. In certain embodiments, the method comprises hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula V1-B-3′ with a second population of oligonucleotides comprising a bottom strand sequence having the following formula V2′-B′-3′ to provide a population of duplexes. After hybridizing, the 3′ ends of the oligonucleotides in the duplexes are extended to produce a population of double stranded products comprising a top strand sequence having the following formula V1-B-V2, where V2 is complementary to V2′.

主权项

1. A method comprising:
a) hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′ with a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′ to provide a population of duplexes; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;the nucleotide sequence of B is the same for each oligonucleotide of said first population;the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;the nucleotide sequence of V1 is variable between the oligonucleotides of the first populationthe nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and V1 and V2′ hybridize to sites in a reference genome; and b) extending the 3′ ends of the oligonucleotides in said duplexes to produce a population of double stranded products comprising a top strand sequence having the following formula:
V1-B-V2, wherein V2 is complementary to V2′.